CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
The wash Buffer is designed as washing solution for immunohistochemical and immunocytological staining procedures on slides. Wash Buffer is primarily used with formalin-fixed paraffin-embedded tissue sections, but also with frozen, HOPE-fixed, and cytological samples as well as in immunoblot procedures. The Wash Buffer is suitable for manually operated and automated immunohistochemical staining.
Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. Washing away the applied reagents after each incubation step is critical to receive optimally stained samples. The Wash Buffer is especially designed for effective washing and therefore ensures brilliant staining results.
Principle of method:
The Wash Buffer is a 20fold concentrated phosphate buffer with additives of sodium chloride, detergent, and stabilising substances. For preparation of the working strength solution the buffer concentrate is diluted 1:20 with deionised or distilled water. The resulting solution has a pH of 7.2 (7.0 to 7.4). The Wash Buffer is wetting the tissue sections with detergent and thus reduces surface tension and improves spreading the reagents on the tissue section, reduces unspecific binding of reagents on the tissue sample, and because of the exact tuned salt concentration effects an excellent preservation of cell morphology.
Reagents provided:
500 ml Wash Buffer (20fold concentrated, adequate for 10 litres ready-to-use wash buffer)
Storage and handling:
The solution should be stored at room temperature. It is stable up to the expiry date indicated on the label if undiluted. Do not use product after the expiry date. The diluted working strength solution is stable for about 1 week depending on the ambient temperature. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
Preparation of the Wash Buffer working strength solution: Dilute Wash Buffer concentrate 1:20 with deionised or distilled water and mix thoroughly. The pH-value should be at 7.2 (7.0 to 7.4). If necessary adjust pH-value with diluted NaOH or HCl solution.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. The Wash Buffer is a 20fold concentrated solution with a mildly acidic pH-value. The correct pHvalue of about 7.0 (+/- 0.2) is achieved after diluting the solution 1:20. Sometimes deionised water has pH-values considerably different from the neutral point (pH 7.0) depending on the preparation method. Experiments have shown that the Wash Buffer can successfully be diluted with deionised or distilled with water in the pH-range of 5.5 up to 9.5. If a detection system with alkaline phosphatase is used please note: larger amounts of wash buffer remaining on the slides can lead to decreasing enzyme activity. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin 300 is used for stabilisation. A material safety data sheet (MSDS) is available upon request.
HIER T-EDTA Buffer pH 9.0 is a solution developed for heat induced epitope retrieval (HIER) in formalin-fixed paraffinembedded tissue sections on slides. This procedure is primarily used in immunohistochemistry.
Immunohistochemical staining procedures consists of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to antigens the epitopes have to be recovered. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values restore structures of the epitopes making them more accessible to specific antibodies. Enzymatic digestion with proteolytic enzymes is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Principle of method:
HIER T-EDTA Buffer pH 9.0 is a 10fold concentrated EDTA solution in Tris buffer with additives of detergent and stabilising substances. For preparation of the working strength solution the buffer concentrate is diluted 1:10 with deionised or distilled water. The resulting solution has a pH of 9.0 (8.8 to 9.2). HIER T-EDTA Buffer pH 9.0 is a very efficient epitope retrieval solution in immunohistochemical staining procedures to be used with primary antibodies of many different specificities. It leads to considerably stronger signals compared with usually used citrate buffer.
Reagents provided:
100 ml HIER T-EDTA Buffer pH 9.0 (10fold concentrated, adequate for 1 litre ready-to-use T-EDTA Buffer)
Storage and handling:
The solution should be stored at 2-8°C. Do not free ze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. The prepared working strength solution is stable for 1 month, if stored at 2-8°C. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
Preparation of the T-EDTA buffer working strength solution: Dilute HIER T-EDTA Buffer concentrate 1:10 with deionised or distilled water and mix thoroughly. The pH-value should be at 9.0 (8.8 to 9.2). If necessary adjust pH-value with diluted NaOH or HCl solution.
Procedure:
HIER T-EDTA Buffer is suitable for various HIER-methods such as steamer, pressure cooker, autoclave, water bath, and microwave oven. Tissue sections used in heat induced epitope retrieval should always be placed on adhesive slides. Epitope retrieval is carried out after dewaxing and rehydration of the sections. Exemplary protocol using steamer: 1. Prepare the working strength solution by diluting the buffer concentrate as described above and transfer to a Coplin jar. Please make sure that there is enough volume to cover the tissue sections on the slides completely. 2. Fill steamer with water according to instruction manual, close lid and start. 3. After 10 minutes place Coplin jar with T-EDTA buffer in the steamer, close the lid and heat the solution for 20 minutes. 4. Place slides with tissue sections into the preheated solution and close the lid. Tissue sections have to be completely covered with T-EDTA buffer solution. 5. Incubate slides 20 40 minutes. The optimal incubation time needs to be elaborated by the operator. 6. After the incubation take the Coplin jar with slides out of steamer and let cool down at room temperature for about 20 minutes. 7. Remove T-EDTA buffer, rinse slides with wash buffer and proceed with immunohistological staining.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent and specimen with eye, skin or mucous membranes. In case of reagent or specimen coming into contact with a sensitive area, wash area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin 300 is used for stabilisation. A material safety data sheet (MSDS) is available upon request.
HIER Citrate Buffer pH 6.0 is a solution developed for heat induced epitope retrieval (HIER) in formalin-fixed paraffin-embedded tissue sections on slides. This procedure is primarily used in immunohistochemistry.
Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to the antigens the epitopes have to be recovered. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values restore structures of the epitopes making them more accessible to specific antibodies. Enzymatic digestion with proteolytic enzymes is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Principle of method:
HIER Citrate Buffer pH 6.0 is a 10fold concentrated citrate buffer solution with additives of stabilising substances. For preparation of the working strength solution the buffer concentrate is diluted 1:10 with deionised or distilled water. The resulting solution has a pH of 6.0 (5.8 to 6.2). HIER Citrate Buffer pH 6.0 is a very efficient epitope retrieval solution in immunohistochemical staining procedures to be used with primary antibodies of many different specificities.
Reagents provided:
100 ml HIER Citrate Buffer pH 6.0 (10fold concentrated, adequate for 1 litre ready-to-use citrate buffer)
Storage and handling:
The solution should be stored at 2-8°C. Do not free ze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. The prepared working strength solution is stable for 1 month, if stored at 2-8°C. A positive and a negat ive control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Reagent preparation:
Preparation of the citrate buffer working strength solution: Dilute HIER Citrate Buffer concentrate 1:10 with deionised or distilled water and mix thoroughly. The pH-value should be at 6.0 (5.8 to 6.2). If necessary adjust pH-value with diluted NaOH or HCl solution.
Procedure:
HIER Citrate Buffer is suitable for various HIER-methods such as steamer, pressure cooker, autoclave, water bath, and microwave oven. Tissue sections used in heat induced epitope retrieval should always be placed on adhesive slides. Epitope retrieval is carried out after dewaxing and rehydration of the sections. Exemplary protocol using steamer: 1. Prepare the working strength solution by diluting the buffer concentrate as described above and transfer to a Coplin jar. Please make sure that there is enough volume to cover the tissue sections on the slides completely. 2. Fill steamer with water according to instruction manual, close lid and start. 3. After 10 minutes place Coplin jar with citrate buffer in the steamer, close the lid and heat the solution for 20 minutes. 4. Place slides with tissue sections into the preheated solution and close the lid. Tissue sections have to be completely covered with citrate buffer solution. 5. Incubate slides 20 40 minutes. The optimal incubation time needs to be elaborated by the operator. 6. After the incubation take the Coplin jar with slides out of steamer and let cool down at room temperature for about 20 minutes. 7. Remove citrate buffer, rinse slides with wash buffer and proceed with immunohistological staining.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin 300 is used for stabilisation. A material safety data sheet (MSDS) is available upon request.
Aqueous Mount is an aqueous mounting medium for histological and cytological preparations. It is especially suitable for mounting of tissue sections stained with alcohol-soluble chromogenic substrates such as Fast Red or AEC. Even sections stained with alcohol-resistant chromogenic substrates can be mounted with Aqueous Mount if they were not previously dehydrated. Aqueous Mount is intended for research use only.
Aqueous Mount in the following formats: 30 ml Ready-to-use
Storage and handling:
The solution should be stored at room temperature in the original container without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
The solution is ready-to-use
Procedure:
Apply a sufficient amount of Aqueous Mount onto the stained histological or cytological section; depending on the size of section 2 to 5 drops. Cover with a coverslip. Heating of the mounted section for hardening is not necessary. Coverslips can be removed by soaking the section over night in water.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. In case of swallowing rinse the mouth excessively with large amounts of water and turn to a doctor. A material safety data sheet (MSDS) is available upon request.
The BrightDiluents universal IHC diluent is specially formulated to stabilize antibodies and eliminate backgrounds encountered in immunohistochemistry staining procedures. The diluent contains complex proteins and non-protein mixtures which eliminate histostaining background by blocking Fc receptors and prevents non-specific protein-protein interactions. It can also be used as blocking applied before primary antibodies. If primary antibodies are diluted in this diluent, a blocking step before primary antibodies can generally be omitted. The blocking /diluent solution is supplied in Ready-to-Use format. It should not be diluted to be effective. The diluent contains no azide, therefore, it can be used to dilute and stabilize HRP-conjugates as well as any other solution that need and stabilizing carrier protein, such as Blocking and Poly-AP conjugates. This product should be interpreted by a qualified pathologist with relevant clinical information, morphological and histological studies and with proper controls. The ready-to-use antibody diluent is available in different colors: MON-APP917 (Transparant/colorless), MON-APP917B (Blue), MON-APP917Y (Yellow), MON-APP917G (green), MON-APP917R (Red)
Principle of method:
Antibody diluent
Reagents provided:
125 ml BrightDiluent, normal antibody diluent (ready-to-use)
The BrightDiluents universal IHC diluent is specially formulated to stabilize antibodies and eliminate backgrounds encountered in immunohistochemistry staining procedures. The diluent contains complex proteins and non-protein mixtures which eliminate histostaining background by blocking Fc receptors and prevents non-specific protein-protein interactions. It can also be used as blocking applied before primary antibodies. If primary antibodies are diluted in this diluent, a blocking step before primary antibodies can generally be omitted. The blocking /diluent solution is supplied in Ready-to-Use format. It should not be diluted to be effective. The diluent contains no azide, therefore, it can be used to dilute and stabilize HRP-conjugates as well as any other solution that need and stabilizing carrier protein, such as Blocking and Poly-AP conjugates. This product should be interpreted by a qualified pathologist with relevant clinical information, morphological and histological studies and with proper controls. The ready-to-use antibody diluent is available in different colors: MON-APP917 (Transparant/colorless), MON-APP917B (Blue), MON-APP917Y (Yellow), MON-APP917G (green), MON-APP917R (Red)
Principle of method:
Antibody diluent
Reagents provided:
125 ml BrightDiluent, normal antibody diluent (ready-to-use)
The BrightDiluents universal IHC diluent is specially formulated to stabilize antibodies and eliminate backgrounds encountered in immunohistochemistry staining procedures. The diluent contains complex proteins and non-protein mixtures which eliminate histostaining background by blocking Fc receptors and prevents non-specific protein-protein interactions. It can also be used as blocking applied before primary antibodies. If primary antibodies are diluted in this diluent, a blocking step before primary antibodies can generally be omitted. The blocking /diluent solution is supplied in Ready-to-Use format. It should not be diluted to be effective. The diluent contains no azide, therefore, it can be used to dilute and stabilize HRP-conjugates as well as any other solution that need and stabilizing carrier protein, such as Blocking and Poly-AP conjugates. This product should be interpreted by a qualified pathologist with relevant clinical information, morphological and histological studies and with proper controls. The ready-to-use antibody diluent is available in different colors: MON-APP917 (Transparant/colorless), MON-APP917B (Blue), MON-APP917Y (Yellow), MON-APP917G (green), MON-APP917R (Red)
Principle of method:
Antibody diluent
Reagents provided:
125 ml BrightDiluent, normal antibody diluent (ready-to-use)
The BrightDiluents universal IHC diluent is specially formulated to stabilize antibodies and eliminate backgrounds encountered in immunohistochemistry staining procedures. The diluent contains complex proteins and non-protein mixtures which eliminate histostaining background by blocking Fc receptors and prevents non-specific protein-protein interactions. It can also be used as blocking applied before primary antibodies. If primary antibodies are diluted in this diluent, a blocking step before primary antibodies can generally be omitted. The blocking /diluent solution is supplied in Ready-to-Use format. It should not be diluted to be effective. The diluent contains no azide, therefore, it can be used to dilute and stabilize HRP-conjugates as well as any other solution that need and stabilizing carrier protein, such as Blocking and Poly-AP conjugates. This product should be interpreted by a qualified pathologist with relevant clinical information, morphological and histological studies and with proper controls. The ready-to-use antibody diluent is available in different colors: MON-APP917 (Transparant/colorless), MON-APP917B (Blue), MON-APP917Y (Yellow), MON-APP917G (green), MON-APP917R (Red)
Principle of method:
Antibody diluent
Reagents provided:
125 ml BrightDiluent, normal antibody diluent (ready-to-use)
The BrightDiluents universal IHC diluent is specially formulated to stabilize antibodies and eliminate backgrounds encountered in immunohistochemistry staining procedures. The diluent contains complex proteins and non-protein mixtures which eliminate histostaining background by blocking Fc receptors and prevents non-specific protein-protein interactions. It can also be used as blocking applied before primary antibodies. If primary antibodies are diluted in this diluent, a blocking step before primary antibodies can generally be omitted. The blocking /diluent solution is supplied in Ready-to-Use format. It should not be diluted to be effective. The diluent contains no azide, therefore, it can be used to dilute and stabilize HRP-conjugates as well as any other solution that need and stabilizing carrier protein, such as Blocking and Poly-AP conjugates. This product should be interpreted by a qualified pathologist with relevant clinical information, morphological and histological studies and with proper controls. The ready-to-use antibody diluent is available in different colors: MON-APP917 (Transparant/colorless), MON-APP917B (Blue), MON-APP917Y (Yellow), MON-APP917G (green), MON-APP917R (Red)
Principle of method:
Antibody diluent
Reagents provided:
125 ml BrightDiluent, normal antibody diluent (ready-to-use)
AEC Single Solution is a ready-to-use solution intended for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). AEC (3-Amino-9-ethylcarbazol) leads to the formation of a red-brown precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous mounting media and can be observed by light microscopy. AEC Single Solution is especially useful when a high sensitivity is desired.
The solution should be stored at 2-8°C without furt her dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. AEC Single Solution is a ready-to-use solution. Preparation of a working solution as in other chromogenic substrates is not necessary. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
The solution is ready-to-use. AEC Single Solution can be used directly from the refrigerator and should be stored again at 2-8°C aft er use. When using the small package (8 ml MON-APP169) please directly drop from the bottle. When using the 100 ml package (MON_APP?) please transfer up to 8 ml of the AEC Single Solution into one of the provided dropper bottles. The transferred solution is stable for many weeks if stored at 2-8°C. The vo lume required for several staining runs should be transferred so that the 100 ml stock bottle has to be opened only a few times. If you would like to pipette the solution use a clean vial from which you pipette. Remaining quantities should not be filled back into the bottle but disposed as hazardous material.
Procedure:
1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply the AEC Single Solution to the slide. Incubate for 3-6 minutes. (Incubation time can be extended up to 30 minutes, if desired.) 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for at least 5 minutes. 7) Mount with an aqueous mounting medium.
Expected results:
During the reaction of the substrate with horse radish peroxidase in presence of the chromogen AEC, a red-brown precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous solvents and can be observed by light microscopy.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, or contact our technical support. Also refer to the instructions of the detection systems for guidance on general troubleshooting.
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex method in which histological as well as immunological detection methods are combined. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). In some tissues endogenous peroxidase activity may cause non-specific staining. The enzyme activity should be blocked by incubation with hydrogen peroxide solution (H2O2 solution). The step is carried out before incubation with primary antibody but after dewaxing and rehydration. Background staining due to endogenous biotin can be blocked through an avidin-biotin blocking step prior to the primary antibody incubation step. The coloured precipitate formed by AEC is soluble in organic solvents. The tissue sections therefore have to be counterstained with aqueous solutions (e. g. Gills or Mayers haematoxylin) and mounted with aqueous mounting media. The colour intensity of the reaction product can decrease with time, especially when exposed to light. The staining reaction itself can be influenced in the same way when carried out in strong light. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided
Precautions:
Use by qualified personnel only. AEC (3-Amino-9-ethylcarbazol) and the solvents used are considered hazardous materials. Material safety data sheets (MSDS) are available upon request. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Oxidising substances, e. g. metals, dust, bacteria or glass devices can influence the stability of AEC Single Solution. Such contaminations have to be avoided. Non-consumed solution needs to be discarded as dangerous substance.
The blocking solution Peroxide Block is intended for inhibition of endogenous peroxidase activity in tissue sections. It is primarily intended to be used in immunohistochemistry on formalin-fixed paraffin-embedded samples if using a detection system with horse radish peroxidase.
Endogenous peroxidase activity in the tissue can result in unspecific background staining in immunohistochemical staining procedures using horse radish peroxidase (HRP) as detection enzyme. This effect can be eliminated when tissue sections are incubated with Peroxide Block prior to immunohistochemical staining. Hydrogen peroxide in the solution blocks the activity of endogenous peroxidase.
Principle of method:
Peroxide Block is applied onto tissue sections to reduce non-specific staining due to endogenous peroxidase activity in immunohistochemistry. The step is carried out before incubation with primary antibody but after dewaxing and rehydration. If a heat induced epitope retrieval (HIER) or enzymatic digestion is necessary for immunohistochemical detection it is of no importance if the Peroxide Block is used before or after this step. In some cases it has been shown, that blocking of endogenous peroxidase before the epitope retrieval leads to better results.
Reagents provided:
8 ml Peroxide Block (ready-to-use)
Storage and handling:
The solution should be stored at 2-8°C without furt her dilution. Please store the reagent in a dark place and do not freeze it. Avoid exposure to strong light. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Procedure:
1. Apply Peroxide Block for 10 minutes at room temperature. The section should be covered completely. 2. Rinse with wash buffer. 3. Proceed with next steps for immunohistochemical staining as usual starting with protein blocking or the primary antibody.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems containing Peroxide Block for guidance on general troubleshooting
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific binding. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. A material safety data sheet (MSDS) is available upon request
Antibody Diluent is developed for dilution of primary antibodies. Antibodies diluted with Antibody Diluent are primarily used in immunohistochemistry with formalin-fixed paraffin-embedded tissue sections, but also with frozen, HOPE-fixed, and cytological samples as well as in immunoblot procedures.
Antibody diluents used in immunohistochemistry should protect the antibody from microbial contamination and stabilize the antibody chemically. Antibody Diluent reduces non-specific binding of antibodies to tissue sections and is therefore extremely useful in receiving background-free staining results.
Principle of method:
Immunohistochemical staining procedures often start with incubation of a blocking solution to reduce unspecific binding of primary antibody to tissue sections. This step can be omitted if the antibody used is diluted in Antibody Diluent. Antibody Diluent ? minimises unspecific binding of the primary antibody to the tissue section, ? reduces surface tension of the antibody solution and improves spreading the reagent on the slide, ? increases microbial and chemical stability of the antibody, ? reduces adhesion of antibody to the surface of the vial, ? and minimises the danger of antibody degradation by proteolytic enzymes.
Reagents provided:
100 ml Antibody Diluent (ready-to-use)
Storage and handling:
The solution should be stored at 2-8°C without further dilution. Do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems containing Peroxide Block for guidance on general troubleshooting
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results. Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use through qualified personnel only. Wear protective clothing to avoid contact of reagents and specimens with eye, skin and mucous membranes. If reagents or specimens come in contact with sensitive area, wash with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. ProClin300 and sodium azide (NaN3) are used for stabilisation. Reaction of sodium azide with lead or copper in drainage pipes can result in the formation of highly explosive metallic azides. Discard the antibody solution in a large volume of running water to avoid formation of deposits. A material safety data sheet (MSDS) for the pure substances is available upon request.
The aqua mounting medium is a widely used for coverslipping from aqueous solutions and is non-fluorescing and has an antifade component to increase the viewing time of the specimen. Use Aqua-Poly/Mount with most fluorescent dyes and stains including DAB, Alkaline Phosphatase, Fast Red, AEC (aminoethylcarbazole) and a variety of other chromogens to enhance and retain fluorescent intensity. The mounting medium (water based) can be used for frozen sections, fat stains, chromogens for immunohistochemistry and in situ hybridization as well as other applications requiring a water-soluble mounting medium.
Principle of method:
Coverslipping from aqueous solutions
Reagents provided:
50 ml Mounting medium (water based), ready-to-use.
Storage and handling:
Store at room temperature
Procedure:
1. Prepare slides as required. 2. Prior to coverslipping rinse the slides in distilled or deionized water. 3. Place the bottle upside down with the end cap attached in a container before use. This will help clear of tiny bubbles. 4. Blot excess water from the slide without drying the tissue specimen. The tissue must be moist prior to mounting. 5. Apply enough drops of aqua mounting medium on the tissue section so that the tissue is completely covered. 6. Carefully lower the coverslip at an angle while gently applying pressure to force any excess medium and air bubbles away from the tissue and out from under the coverslip. 7. Slides can be dried at room temperature or at 4°C or in an oven but the temperature hight is depending what type of staining is used. 8. Allow the aqua mounting medium to dry before examine under the microscope.
Aqueous Mount is an aqueous mounting medium for histological and cytological preparations. It is especially suitable for mounting of tissue sections stained with alcohol-soluble chromogenic substrates such as Fast Red or AEC. Even sections stained with alcohol-resistant chromogenic substrates can be mounted with Aqueous Mount if they were not previously dehydrated. Aqueous Mount is intended for research use only.
Aqueous Mount in the following formats: 60 ml Ready-to-use
Storage and handling:
The solution should be stored at room temperature in the original container without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.
Reagent preparation:
The solution is ready-to-use
Procedure:
Apply a sufficient amount of Aqueous Mount onto the stained histological or cytological section; depending on the size of section 2 to 5 drops. Cover with a coverslip. Heating of the mounted section for hardening is not necessary. Coverslips can be removed by soaking the section over night in water.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagents or specimen with eye, skin or mucous membrane. In case of a reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. In case of swallowing rinse the mouth excessively with large amounts of water and turn to a doctor. A material safety data sheet (MSDS) is available upon request.
Antibody Diluent B is especially developed for dilution of certain primary antibodies. Antibodies diluted with Antibody Diluent B are primarily used in immunohistochemistry with formalin-fixed paraffin-embedded tissue sections, but also with frozen, HOPE-fixed, and cytological samples as well as in immunoblot procedures.
Antibody diluents used in immunohistochemistry should protect the antibody from microbial contamination and stabilize the antibody chemically. Antibody Diluent B reduces non-specific binding of antibodies to tissue sections and is therefore extremely useful in receiving background-free staining results.
Principle of method:
Immunohistochemical staining procedures often start with incubation of a blocking solution to reduce unspecific binding of primary antibody to tissue sections. This step can be omitted if the antibody used is diluted in Antibody Diluent B. Antibody Diluent B minimises unspecific binding of the primary antibody to the tissue section, reduces surface tension of the antibody solution and improves spreading the reagent on the slide, increases microbial and chemical stability of the antibody, reduces adhesion of antibody to the surface of the vial, and minimises the danger of antibody degradation by proteolytic enzymes.
Reagents provided:
25 ml Antibody Diluent B (ready-to-use)
Storage and handling:
The solution should be stored at 2-8°C without furt her dilution. Do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date. If stored at room temperature the solution is stable for at least 10 month from the date of delivery. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by this reagent, please contact our technical support.
Expected results:
During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Trouble shooting:
If you observe unusual staining or other deviations from the expected results please read these instructions carefully, contact our technical support. Also refer to the instructions of the detection systems containing Peroxide Block for guidance on general troubleshooting
Quality Control:
We recommend carrying out a positive and a negative control with every staining run. The positive control permits the validation of appropriate processing of the sample. If the negative control has a positive result, this points to unspecific staining. Please refer to the instructions of the detection system for guidance on general quality control procedures.
Performance characteristics:
Studies have been conducted to evaluate the performance of the kit reagents. The product has been found to be suitable for the intended use
Limitations of procedure:
Immunohistochemistry is a complex technique involving both histological and immunological detection methods. It requires a highly trained histotechnologist. Tissue processing and handling prior to immunostaining, for example variations in fixation and embedding or the inherent nature of the tissue can cause inconsistent results (Nadji and Morales, 1983). Inadequate counterstaining and mounting can influence the interpretation of the results.Sanbio guarantees that the product will meet all requirements described from its shipping date until its expiry date, as long as the product is correctly stored and utilized. No additional guarantees can be given. Under no circumstances shall Sanbio be liable for any damages arising out of the use of the reagent provided.
Precautions:
Use by qualified personnel only. Wear protective clothing to avoid contact of reagent or specimen with eye, skin or mucous membrane. In case of the reagent or specimen coming into contact with a sensitive area, wash the area with large amounts of water. Microbial contamination of the reagent must be avoided, since otherwise non-specific staining may occur. (NaN3), used for stabilisation, is not considered hazardous material in the concentration used. Sodium azide deposits in drainage pipes made of lead or copper can result in the formation of highly explosive metallic azides. To avoid such deposits in drainage pipes, sodium azide should be discarded in a large volume of running water. A material safety data sheet (MSDS) is available upon request.
Paraformaldehyde (PFA) is the polymerization product of formaldehyde with a typical degree of polymerization of 8100 units. Paraformaldehyde is not a fixative itself; it must be depolymerized to formaldehyde in solution. In cell culture, a typical formaldehyde fixing procedure would involve using a 4% formaldehyde solution in phosphate buffered saline (PBS) on ice for 10 minutes. Fixing ensures that sample cell structures stay intact and that antigens are immobilized, while ideally still permitting unfettered access of antibodies to target antigens. CAS 30525-89-4, soluble in water, pH 7.0-7.6 at 25°C. The solution should be clear, colorless, with no precipitate.
Principle of method:
Fixing cells for immunohistochemistry (IHC)
Reagents provided:
500 ml 4 % Paraformaldehyde (PFA) solution in PBS
Storage and handling:
Store at -20°C for one year
Trouble shooting:
Adjust pH to 7.2-7.6 with NaH2PO4 if pH
Precautions:
Causes serious eye damage. Suspected of causing cancer. May cause an allergic skin reaction. Causes skin irritation. Avoid breathing dust/fume/gas/mist/vapors/spray. Wear protective gloves/protective clothing/eye protection/face protection. Use personal protective equipment as required. IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. Store locked up. Dispose of contents/container in accordance with local/regional/national/international regulations
The Citrate buffer, is designed for use during heat-induced epitope retrieval (HIER) on formalinfixed paraffin-embedded tissue sections prior to antibody application. In immunohistochemistry (IHC), protein cross-links are formed during formalin or other aldehyde fixation. These cross-links mask the antigenic sites in tissue specimens and results in weak or false negative staining in immunohistochemical detection of proteins. The use of this citrate buffer in combination with heat ensures that the antigenicity of proteins modified, thus enhancing staining intensity of antibodies. For the recommended pre-treatment technique, please refer to the individual antibody's instructions for use. This buffer is a 10x concentrate solution and must be diluted 1:10 with deionized water before using. The final readyto-use solution should have a pH of 6.0 ± 0.3. The buffer can be used for a maximum of three runs within five days after dilution. Also available in 250 ml, 500 ml en 1000 ml.
Principle of method:
Heat-Induced Epitope Retrieval
Reagents provided:
100 ml 10x concentrate solution. Citrate buffer, Heat-Induced Epitope Retrieval (10x, Tween20), Red
Storage and handling:
Store at room temperature
Reagent preparation:
Before use dilute 1:10 with deionized water. The final ready-to-use solution should have a pH of 6.0 ± 0,3,
Procedure:
1. Deparaffinize and rehydrate the tissue slides. 2. Fill the PT Module tank with 150 ml of citrate buffer and 1,350 ml of deionized water, to obtain 1,500 ml of ready-to-use solution (1:10 dilution). 3. Pre-heat the PT Module until temperature reaches 65°C and place the formalin-fixed slides in slide racks into the PT Module tank. 4. Heat the citrate buffer and slides to 98°C and incubate for 20 minutes. 5. Cool slides in the PT Module to 65°C. 6. Remove slides and cool to room temperature for at least 5 minutes in a PBS or TBS based buffer. 7. Continue with staining according to IHC protocol. Note: Alternative heating sources, such as a microwave or a pressure cooker, can be used to replace the PT Module tank. The optimal incubation time for these heating sources should be determined by authorised personnel / researchers.
The TRIS/EDTA buffer, is designed for use during heat-induced epitope retrieval (HIER) on formalinfixed paraffin-embedded tissue sections prior to antibody application. In immunohistochemistry (IHC), protein cross-links are formed during formalin or other aldehyde fixation. These cross-links mask the antigenic sites in tissue specimens and results in weak or false negative staining in immunohistochemical detection of proteins. The use of this citrate buffer in combination with heat ensures that the antigenicity of proteins modified, thus enhancing staining intensity of antibodies. For the recommended pre-treatment technique, please refer to the individual antibody's instructions for use. This buffer is a 10x concentrate solution and must be diluted 1:10 with deionized water before using. The final readyto-use solution should have a pH of 9.0 ± 0.3. The buffer can be used for a maximum of three runs within five days after dilution. Also available in 250 ml, 500 ml en 1000 ml.
Principle of method:
Heat-Induced Epitope Retrieval
Reagents provided:
100 ml 10x concentrate solution. TRIS/EDTA buffer, Heat-Induced Epitope Retrieval (10x, Tween20), Blue
Storage and handling:
Store at room temperature
Reagent preparation:
Before use dilute 1:10 with deionized water. The final ready-to-use solution should have a pH of 9.0 ± 0,3,
Procedure:
1. Deparaffinize and rehydrate the tissue slides. 2. Fill the PT Module tank with 150 ml of TRIS?EDTA buffer and 1,350 ml of deionized water, to obtain 1,500 ml of ready-to-use solution (1:10 dilution). 3. Pre-heat the PT Module until temperature reaches 65°C and place the formalin-fixed slides in slide racks into the PT Module tank. 4. Heat the TRIS/EDTA buffer and slides to 98°C and incubate for 15 minutes. 5. Cool slides in the PT Module to 65°C. 6. Remove slides and cool to room temperature for at least 5 minutes in a PBS or TBS based buffer. 7. Continue with staining according to IHC protocol. Note: Alternative heating sources, such as a microwave or a pressure cooker, can be used to replace the PT Module tank. The optimal incubation time for these heating sources should be determined by authorised personnel / researchers.
The TRIS/EDTA buffer, is designed for use during heat-induced epitope retrieval (HIER) on formalinfixed paraffin-embedded tissue sections prior to antibody application. In immunohistochemistry (IHC), protein cross-links are formed during formalin or other aldehyde fixation. These cross-links mask the antigenic sites in tissue specimens and results in weak or false negative staining in immunohistochemical detection of proteins. The use of this citrate buffer in combination with heat ensures that the antigenicity of proteins modified, thus enhancing staining intensity of antibodies. For the recommended pre-treatment technique, please refer to the individual antibody's instructions for use. This buffer is a 10x concentrate solution and must be diluted 1:10 with deionized water before using. The final readyto-use solution should have a pH of 9.0 ± 0.3. The buffer can be used for a maximum of three runs within five days after dilution. Also available in 250 ml, 500 ml en 1000 ml.
Before use dilute 1:10 with deionized water. The final ready-to-use solution should have a pH of 9.0 ± 0,3,
Procedure:
1. Deparaffinize and rehydrate the tissue slides. 2. Fill the PT Module tank with 150 ml of TRIS?EDTA buffer and 1,350 ml of deionized water, to obtain 1,500 ml of ready-to-use solution (1:10 dilution). 3. Pre-heat the PT Module until temperature reaches 65°C and place the formalin-fixed slides in slide racks into the PT Module tank. 4. Heat the TRIS/EDTA buffer and slides to 98°C and incubate for 15 minutes. 5. Cool slides in the PT Module to 65°C. 6. Remove slides and cool to room temperature for at least 5 minutes in a PBS or TBS based buffer. 7. Continue with staining according to IHC protocol. Note: Alternative heating sources, such as a microwave or a pressure cooker, can be used to replace the PT Module tank. The optimal incubation time for these heating sources should be determined by authorised personnel / researchers.
Expected results:
Epitope retrieval
Precautions:
Refer to SDS
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