For in-vitro Diagnostic Use. The BrightVision one step detection system Goat Anti-Rabbit IgG HRP and Goat anti-Mouse IgG AP, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision detection system, peroxidase Goat Anti-Rabbit HRP and Goat anti-Mouse AP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Rabbit and Mouse bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen. The clinical interpretation of any staining or its absence should be determined by a qualified pathologist and complemented by morphologic studies; controls should be evaluated within the context of the patients clinical history and/or other diagnostic tests.
Principle of method:
BrightVision, one step detection system Goat Anti- Rabbit HRP and Mouse AP (Ready-to-Use).
Reagents provided:
BrightVision, One step detection system Goat anti-Rabbit HRP / Goat anti-Mouse AP (Ready-to-use; 55 ml)
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse and/or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9 Detection system, polymer Rabbit HRP/Mouse AP, (Labeled polymer; 30 min), 10. Wash buffer (TBS buffer; 2x 5 min), 11. Substrate (DAB; see applicble IFU), 12. Wash aqua dest (Wash; 2x 2 min), 13. Substrate (Fast Red / New Fuchsin; see applicable IFU), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain and coverslip with aqueous mounting medium.
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
The BrightDAB substrate DAB (3,3Diaminobenzidine) is a widely used chromogen for immunohistochemical staining and immunoblotting. When in the presence of peroxidase enzyme, DAB produces a brown precipitate that is insoluble in alcohol and xylene. This product comes in a two-component system consisting of a liquid stable DAB chromogen and DAB substrate buffer. DAB Solution A: Readyto-Use Buffered H2O2,; DAB Solution B: Concentrated DAB solution. BrightDAB is a very stable and superior formulation of DAB. In some cases, antibodies titers may increase by two-fold. BrightDAB can be used both manually and on automated stainers. The clinical interpretation of any staining or its absence should be determined by a qualified pathologist and complemented by morphologic studies; controls should be evaluated within the context of the patients clinical history and/or other diagnostic tests. Also available in 500 ml and 1000 ml.
Principle of method:
DAB substrate for use with HRP-labeled detection systems
Reagents provided:
DAB Solution A: Buffered H2O2 (Ready-to-Use) 110 ml DAB Solution B: Concentrated DAB solution 5 ml
Storage and handling:
2-8°C and in the dark
Reagent preparation:
1 Working solution: Add 40 ?l DAB Solution B (± one drop) to 1 ml Solution A, mix well. 2 Incubate the DAB solution one time 8 minutes without washing in between.
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, polymer HRP, (Labeled polymer; 30 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Substrate (DAB; 8 min), 12. Wash aqua dest (Wash; 2x 2 min), 13. Counterstain, dehydrate and coverslip (Auxiliary)
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
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)
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.
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 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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