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 dilution is 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. The predicted molecular weight of serotonin transporter based on its nucleic acid sequence is 67,815. The actual weight that one might observe from tissue extracts could be higher due to glycosylation. We validated the antibody with rat SERT overexpressed in an insect cell expression system. The MW from that was about 67 kD. Our SERT antibody dilution was 1:20,000 using chemiluminescent detection.
Immunolabeling is completely abolished by pre-adsorption with synthetic rat 5HT transporter (602-622).
照片描述: IHC image of neurons staining for the 5-HT transporter in the raphe nucleus of the rat brainstem (above). The tissue was fixed with 4% formaldehyde in 0.1 M phosphate buffer, before being removed and prepared for vibratome sectioning. Floating sections were incubated at RT in 10% goat serum in PBS, before standard IHC procedure. Primary antibody was incubated at 1:10000 for 48 hours, goat anti-rabbit secondary was subsequently added for 1 hour after washing with PBS. Light microscopy staining was achieved with standard biotin-streptavidin/HRP procedure and DAB chromogen. Florescent secondary (below)
规格/体积: 100 µL
性状: Lyophilized Whole Serum
反应性: Human, Marine, Monkey, Mouse, Rat, Sparrow
别名: 5HTT; HTT; Sodium-dependent serotonin transporter; Solute carrier family 6 member 4; Slc6a4; solute carrier family 6 (neurotransmitter transporter, serotonin), member 4
免疫原: Rat 5-HT transporter (579-599)
Gene Symbol: Slc6a4