The 5-HIAA antibody was raised to 5-HIAA coupled to BSA with paraformaldehyde. The antibody produces moderate labeling of raphe neurons in normal rat. In rats whose serotonergic system has been activated, staining intensity is increased to a maximum label. Recommended dilutions of the antiserum are 1/200-1/400 for indirect immunofluorescence and 1/4000-1/8000 for biotin-streptavidin/HRP technique. The specificity of the antiserum was evaluated using a model system of gelatin-indole plugs by a method similar to published procedures (Schipper and Tilders, 1983). Results showed that the 5-HIAA antibody dose dependently stained 5-HIAA but did not stain any concentration of 5-HT or 5-HTP. The antiserum was also tested by pre-adsorption at 25 µg/mL with various BSA conjugates. While pre-adsorption with 5-HIAA conjugate completely eliminates immunolabeling, pre-adsorption with conjugates of 5-HT,5-HTP and dopamine had no effect on staining intensity or distribution of stain.
The 5-HT Antibody was raised in rabbit against 5-HTP coupled to BSA with paraformaldehyde. The antibody has a proven maximum biotin-streptavidin/HRP staining at a 1/1000 - 1/2000 dilution in rat raphe nuclei. Optimal dilution will vary depending upon fixation, labeling technique and/or detection system; therefore, a dilution series is recommended. The specificity of the antiserum was evaluated using a model system of gelatin-indole plugs by a method similar to published procedures (Shipper and Tilders, 1983). Results showed that the 5-HTP antibody dose dependently stained 5-HTP but did not stain any concentration of 5-HT or 5-HIAA. The antiserum was also tested by pre-adsorption with indole/paraformaldehyde/BSA conjugates. Staining was completely blocked by pre-adsorption with 5-HTP conjugate and unaffected by 5-HIAA or 5-HT conjugate.
The 5-HT 1A Receptor Antibody was raised against synthetic peptide sequence corresponding to amino acids 294-312 of the rat 5-HT1A receptor. The antiserum is provided as 100 µL of affinity purified serum containing 1% BSA. The antiserum demonstrates strongly positive labeling of rat cortex, arcuate and hippocampus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/100 - 1/200 in PBS - Bn/Av-HRP detection. Intensification methods such as nickel will approximately double the dilution factor as recommended. The antibody was characterized by immunohistochemistry and Western blot. Western blot showed a single band of approximately 45 kD. Preincubation of the antibody with an excess of the synthetic peptide blocked staining. Immunohistochemical staining of rat brain correlates well with Northern analysis, in situ hybridization and receptor autoradiography. BlastP database sequence homology searches confirmed that this sequence is unique to rat, mouse and human 5-HT1A receptors.
The 5-HT 2A Receptor Antibody was raised against a multiple antigenic peptide of an N-terminal synthetic sequence corresponding to amino acids 22-41 of rat 5HT2A receptor. The antibody is provided as 100 uL of affinity purified serum in PBS (0.02 M sodium phosphate with 0.15 M sodium chloride, pH 7.5) with 1% BSA (bovine serum albumin), and 0.02% sodium azide. The antiserum demonstrates strongly positive labeling of rat cortex, amygdala and hippocampus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/300 - 1/500 in PBS/0.03% Triton X-100 - Bn/Av-HRP Technique. The addition of intensifying reagents such as nickel ammonium sulfate to the chromogen solution will approximately double the dilution factor as recommended. Immunolabeling is completely abolished by preadsorption with synthetic rat 5HT2A receptor (22-41). Immunolabeling of Western blot revealed a single band of approximately 53kD.
The ImmunoStar peptide control for 5-HT2A Receptor is intended for the immuno-adsorption of 5-HT2A Receptor antiserum, catalog number 24288. Pre-adsorption of 5-HT2A Receptor antiserum, diluted according to the antibody specification sheet, with 5 µg/ml 5-HT2A Receptor peptide immunogen following the instructions below provides complete blockage of 5-HT2A Receptor immunolabeling. The peptide is provided as 25 µg of lyophilized rat 5-HT2A Receptor, sequence 22-41. Also, this antiserum contains 0.09% sodium azide. Please read the instructions carefully before beginning the procedure.
The 5-HT 2C Receptor Antibody was raised against synthetic peptide sequence corresponding to amino acids 439-460 of the rat 5-HT2C receptor coupled to KLH and bovine thyroglobulin. The ImmunoStar 5-HT2C receptor antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat choroid plexus and hippocampus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are /500 - 1/1000 in PBS - Bn/Av-HRP technique. Intensification methods such as nickel will approximately double the dilution factor as recommended. The antibody was characterized by immunohistochemistry and Western blot. Western blotting revealed a single band of approximately 70 kD. Preincubation of the antibody with an excess of the synthetic peptide blocked staining. Immunohistochemical staining of rat brain correlates well with Northern analysis, in situ hybridization and receptor autoradiography. BlastP database sequence homology searches confirmed that this sequence is unique to rat, mouse and human 5-HT2C receptors.
Raised against synthetic peptide sequence corresponding to amino acids 17-34 of the rat 5-HT5A receptor coupled to carrier protein with glutaraldehyde.
Product Type:
Antibody - Antibodies
Antibody Type:
polyclonal
Format:
Liquid
Host Animal:
Rabbit
Species Reactivity:
Clam, Rat
Immunogen:
Rat 5-HT5A receptor (17-34)
Applications:
Immunohistochemistry, Immunocytochemistry, immunofluorescence, Western Blot
The 5-HT 5A Receptor Antibody was raised against synthetic peptide sequence corresponding to amino acids 17-34 of the rat 5-HT5A receptor coupled to carrier protein with glutaraldehyde. The ImmunoStar 5-HT5A Receptor was quality control tested using standard immunohisto-chemical methods. The antiserum demonstrates strongly positive labeling of rat cortex and hippocampus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/100 - 1/300 in PBS/0.3% Triton X-100Â - Cy3 and 1/200Â - 1/400 in PBS/0.3% Triton X-100Â - Bn/Av-HRP. Intensification methods such as nickel will approximately double the dilution factor as recommended. The antibody was characterized by immunoblotting and immunohistochemistry. Immunoblots of rat brain extracts revealed the presence of two bands at molecular weights of 41 and 47 kD. The lower weight band agrees with the calculated molecular weight based on amino acid sequence. The higher weight may represent glycosylated receptor protein. Immunohistochemical staining of rat brain correlates well with Northern blot analysis and in situ hybridization studies. Immunolabeling is completely abolished by preadsorption with synthetic rat 5-HT5A receptor (17-34). BlastP database sequence homology searches indicate that the amino acid sequence is unique to rat and mouse 5-HT5A receptor.
The ImmunoStar 5HT 6-receptor antibody was quality control tested using standard immunohistochemical methods. The antiserum demonstrates significant labeling of rat cortex, amygdala and hippocampus and other areas using indirect immunofluorescent and biotin/avidin-HRP techniques. The addition of intensifying reagents such as nickel ammonium sulfate to the chromogen solution will approximately double the dilution factor as recommended. Immunolabeling is completely abolished by preadsorption with synthetic rat 5HT6 receptor (CLERPPGTPRHPPGPPLW). Immunolabeling of western blot revealed a single band of approximately 53kD.
The 5-HT7 Receptor Antibody was raised against synthetic peptide sequence corresponding to amino acids 8-23 of the rat 5-HT7 receptor coupled to carrier protein with glutaraldehyde. The ImmunoStar 5-HT7 receptor antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat cortex and hippocampus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/100 - 1/300 in PBS - biotin/avidin-HRP Technique. Note: use of Triton X-100 or other detergents is not recommended. The antibody was characterized by immunohistochemistry. Immunohistochemical staining of rat brain correlates well with Northern blot analysis, in situ hybridization and receptor autoradiography studies. Immunolabeling is completely abolished by preadsorption with synthetic rat 5-HT7 receptor (8-23). BlastP database sequence homology searches indicate that the amino acid sequence is unique to rat 5-HT7A, 5-HT7B and 5-HT7C. There is also significant sequence overlap with the mouse and human forms.
The 5-HT BSA/Conjugate Control was prepared by cross-linking 5-HT creatinine sulfate complex to BSA with paraformaldehyde. Pre-adsorption of Serotonin antisera, diluted according to the antibody specification sheet, with 20 µg/ml Serotonin/BSA conjugate following the instructions below provides complete blockage of Serotonin immunolabeling. The conjugate is provided as 50 µg of lyophilized Serotonin creatinine sulfate coupled to BSA with paraformaldehyde.
The 5-HT Goat Rabbit Antibody was raised against serotonin coupled to BSA with paraformaldehyde. The ImmunoStar serotonin antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat hypothalamus and spinal cord using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/400 - 1/800 in PBS/0.3% Triton X-100 - Cy3 Technique and 1/5000 - 1/10,000 in PBS/0.3% Triton X-100 - Bn/Av-HRP Technique. Staining is completely eliminated by pretreatment of the diluted antibody with 100 µg of serotonin/BSA conjugate.
The 5-HT Rabbit Antibody was raised against serotonin coupled to BSA with paraformaldehyde. The ImmunoStar serotonin antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat hypothalamus, raphe nuclei and spinal cord using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/1,000-1/2,000 in PBS/0.3% Triton X-100 - Cy3 Technique and 1/20,000-1/40,000 in PBS/0.3% Triton X-100 - biotin/avidin-HRP Technique. Staining is completely eliminated by pretreatment of the diluted antibody with 25 ug of serotonin/BSA. Cross reactivity of Serotonin antisera was examined. With 5µg, 10µg and 25µg amounts the following substances did not react with Serotonin antisera diluted 1/20,000 using the Bn-SA/HRP labeling method: 5-hydroxytryptophan, 5-hydroxyindole -3- acetic acid, and dopamine.
The 5-HT Transporter was raised to a synthetic peptide corresponding to amino acids 579-599 of rat 5HT transporter coupled to KLH. The ImmunoStar serotonin (5HT) transporter was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat raphe nuclei, hypothalamus, cortex and spinal cord using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions for these methods are 1/800 - 1/1,000 in PBS/0.3% Triton X-100 - Cy3 and 1/10,000 - 1/15,000 in PBS/0.3% Triton X-100 - Biotin/avidin-HRP. By Western blot analysis using rat brain extracts of cortex, hypothalamus, midbrain, and hindbrain, the antibody specifically labels a single band. Immunolabeling is completely abolished by pre-adsorption with synthetic rat 5HT transporter (602-622).
The peptide control for 5-HT Transporter is intended for the immuno-adsorption of 5-HT Transporter antiserum, catalog number 24330. Pre-adsorption of 5-HT Transporter antiserum, diluted according to the antibody specification sheet, with 5 µg/ml 5-HT Transporter peptide immunogen following the instructions below provides complete blockage of 5-HT Transporter immunolabeling. The peptide is provided as 25 µg of lyophilized rat 5-HT Transporter, sequence 602-622. Please read the instructions carefully before beginning the procedure.
The 5-HT Transporter Peptide Control: rat 5-HT Transporter, sequence 579-599. Pre-adsorption of 5-HT Transporter antiserum, diluted according to the antibody specification sheet, with 5 µg/ml 5-HT Transporter peptide immunogen following the instructions below provides complete blockage of 5-HT Transporter immunolabeling. The peptide is provided as 25 µg of lyophilized rat 5-HT Transporter, sequence 602-622.
The ACTH Antibody was raised to ACTH (1-39) purified from porcine pituitary. The antibody produces a maximum fluorescein staining at a 1/100 - 1/200 dilution and a 4+ biotin-streptavidin/HRP staining at a 1/500 - 1/1000 dilution in rat anterior/intermediate pituitary. Optimal dilution will vary depending upon fixation, labeling technique and/or detection system; therefore, a dilution series is recommended. The recommened dilution series are 1/100-1/200 in PBS/0.3% Triton X-100 - FITC Technique and 1/500-1/1000 in PBS/0.3% Triton X-100 - Bn-SA/HRP Technique. Staining is completely eliminated by pretreatment of the diluted antibody with 100 µg/mL of ACTH.
The Alpha-MSH Antibody was raised to synthetic human a-MSH coupled to bovine thyroglobulin with glutaraldehyde. The ImmunoStar alpha melanocyte stimulating hormone antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat pituitary using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/100-1/200 in PBS/0.3% Triton X-100 - FITC Technique and 1/4000-1/6000 in PBS/0.3% Triton X-100 - Bn/Av-HRP Technique. Staining is completely eliminated by pretreatment of the diluted antibody with 100 ug/mL of alpha-MSH.
The Beta-Endorphin Antibody was raised to synthetic human beta endorphin coupled to KLH with carbodiimide. The antibody produces a strong indirect immunofluorescent staining at a 1/200 - 1/400 dilution and a 4+ biotin-streptavidin/HRP staining at a 1/1000 - 1/2000 dilution in rat anterior pituitary. Staining is completely eliminated by pretreatment of the diluted antibody with 10-6 M of Ã-Endorphin. Pre-adsorption of the diluted antibody with 10-6M of the following substances had no effect on -Endorphin labeling: methionine enkephalin, leucine enkaphalin, dynorphin A, dynorphin B, gamma-endorphin, alpha-endorphin, ACTH and alpha-melanocyte stimulating hormone.
The Bombesin Antibody was raised to synthetic human bombesin coupled to bovine thyroglobulin with glutaraldehyde. The ImmunoStar bombesin antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat dorsal horn of spinal cord using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/400-1/600 in PBS/0.3% Triton X-100 - Cy3 Technique and 1/1,000-1/2,000 in PBS/0.3% Triton X-100 - Bn/Av-HRP Technique. Staining is completely eliminated by pretreatment of 1 mL of diluted antibody with 50 µg of bombesin.
The Calbindin D-28K Antibody was raised to Calbindin D-28k purified from bovine cerebellum. The antibody has a proven maximum biotin-streptavidin/HRP staining at a 1/5,000 - 1/10,000 dilution and a 4+ indirect immunofluorescence staining at a 1/500 - 1/1,000 dilution in rat striatum, cortex, and hippocampus. The antiserum has been characterized as specific to calbindin D-28k; please see reference listed below. Optimal dilution will vary depending upon fixation, labeling technique and/or detection system; therefore, a dilution series is recommended.
The Calretinin Antibody was raised to chick calretinin fusion protein. The antibody has a proven maximum biotin-avidin/HRP staining at a 1/2000 - 1/4000 dilution in rat cortex, hippocampus and hypothalamus.
The CCK-8 Antibody was raised to sulphated CCK-8 (26-33) coupled to bovine thyroglobulin with glutaraldehyde. The antibody has a proven strong indirect immunofluorescent staining at a 1/100-1/200 dilution and a strong Biotin-Streptavidin/HRP immunostaining at a 1/500-1/1000 dilution in rat hypothalamus and spinal cord. The specificity of the antiserum was examined by soluble pre-adsorption with the peptides in question at a final concentration of 10-6M CCK-8 immunolabeling was completely abolished by pre-adsorption with CCK-8, gastrin 17 and gastrin 34. Pre-adsorption with the following peptides resulted in no reduction of immunostaining: α-CGRP, -CGRP, neurotensin, somatostatin, substance P, leucine enkephalin, methionine enkephalin, VIP, neuropeptide Y, gastric inhibitory polypeptide, bombesin, glucagon, peptide YY, and FMRF amide.
The C-FOS Antibody was raised to synthetic peptide sequence corresponding to human c-fos (4-17) coupled to bovine thyroglobulin with glutaraldehyde. For induction of c-fos protein activity rats were injected with 1.0 ml of 1.5 M NaCl per 100 grams of body weight. Negative control rats were injected with the same volume of normal saline. The ImmunoStar c-fos antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat paraventricular nucleus and supraoptic nucleus using indirect immunofluorescent and biotin/avidin-HRP techniques. No labeling was seen in negative control rats. Recommended primary dilutions for these methods are 1/200-1/400 in PBS/0.3% Triton X-100 - FITC Technique and 1/4000-1/6000 in PBS/0.3% Triton X-100 - biotin/avidin-HRP Technique. Specificity of the antiserum was demonstrated by blockage of staining in experimental rats by omission of c-fos antibody or by substitution of antibody pre-incubated with synthetic peptide or the conjugate. Immunoblot analysis of mediobasal hypothalamus showed a single band of approximately 55-60 kD.
The peptide control for C-FOS is intended for the immuno-adsorption of C-FOS antiserum, catalog number 26209. Pre-adsorption of C-FOS antiserum, diluted according to the antibody specification sheet, with 5ug/ml C-FOS peptide immunogen following the instructions below provides complete blockage of C-FOS immunolabeling. The peptide is provided as 25ug of lyophilized human C-FOS, and approximately 0.09% sodium azide, sequence 4-17. Please read the instructions carefully.
The CGRP Antibody was raised to rat alpha-CGRP coupled to bovine thyroglobulin with glutaraldehyde. The antibody has a proven fluorescein staining at a 1/100-1/200 dilution and a strong Biotin-Streptadvidin/HRP staining at a 1/2000-1/4000 dilution in rat amygdala, and spinal cord. The specificity of the antiserum was evaluated by soluble pre-adsorption with the peptides in question at a final concentration of 10-5M. CGRP immunolabeling was completely abolished by pre-adsorption with rat α -CGRP and partially eliminated by pre-adsorption with rat -CGRP. Pre-adsorption with the following peptides resulted in no loss of immunostaining: rat amylin, rat adrenomedulin, calcitonin, neurotensin, somatostatin, substance P, leucine enkephalin, methionine enkephalin, VIP, CCK-8, vasopressin and neuropeptide Y.
The CRF Antibody was raised to synthetic ovine CRF. The ImmunoStar serotonin antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat median eminence using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/200 - 1/400 in PBS/0.3% Triton X-100 - Cy3 Technique and 1/1000 - 1/2000 in PBS/0.3% Triton X-100 - Biotin/Avidin-HRP Technique. Immuno-labeling is completely abolished by preadsorption with synthetic CRF at 100 ug per mL of diluted antibody.
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