Light microscopy staining was achieved with standard biotin-streptavidin/HRP procedure and DAB chromogen. Primary antibody was incubated at 1:10000 for 48 hours, goat anti-rabbit secondary was subsequently added for 1 hour after washing with PBS. Floating sections were incubated at RT in 10% goat serum in PBS, before standard IHC procedure. The APOE and Serotonin Transporter Alleles data product (Early, Version 1.0) includes data for the APOE isoform, directly genotyped using a Taqman allelic discrimination SNP assay, where available, or imputed from preexisting genotype array data otherwise, This file also includes human serotonin transporter (5HTTLPR) short and long alleles. The tissue was fixed with 4% formaldehyde in 0.1 M phosphate buffer, before being removed and prepared for vibratome sectioning. Serotonin is a neurotransmitter responsible for regulating mood, digestion, sleep, and more. Pho to Description: IHC image of neurons staining for the 5-HT transporter in the raphe nucleus of the rat brainstem (above). Immunolabeling is completely abolished by pre-adsorption with synthetic rat 5HT transporter (602-622). Our SERT antibody dilution was 1:20,000 using chemiluminescent detection. We validated the antibody with rat SERT overexpressed in an insect cell expression system. The actual weight that one might observe from tissue extracts could be higher due to glycosylation. The predicted molecular weight of serotonin transporter based on its nucleic acid sequence is 67,815. Recommended primary dilution is 1/10,000 – 1/15,000 in PBS/0.3% Triton X-100 – Biotin/avidin-HRP.īy Western blot analysis using rat brain extracts of cortex, hypothalamus, midbrain, and hindbrain, the antibody specifically labels a single band. However, most studies have focused on a single polymorphism (5-HTTLPR) in SLC6A4. The antiserum demonstrates strongly positive labeling of rat raphe nuclei, hypothalamus, cortex and spinal cord using indirect immunofluorescent and biotin/avidin-HRP techniques. The serotoninergic system has been implicated in mood and appetite regulation, and the serotonin transporter gene (SLC6A4) is a commonly studied candidate gene for eating disorders. The ImmunoStar serotonin (5HT) transporter was quality control tested using standard immunohistochemical methods.
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