5-HT (Serotonin) / BSA Conjugate Control

I used the 5HT-BSA conjugate to run a pre-adsorption control for the rabbit anti-serotonin antibody (#20080). 40mg/ml resulted in only very weak labeling of 1:4,000 and 1:10,000 dilutions of the rabbit anti-serotonin antibody, while all labeling was eliminated for dilutions of 1:20,000 and lower. Dilutions of 1:4,000 to 1:80,000 all provided bright labeling in positive controls in which the BSA-5HT conjugate was omitted. The application was immunocytochemistry of sectioned brain tissue from the bumble bee Bombus impatiens (see Paulk et al 2009, J. Comp. Neurol.). DAY 1 Incubate 40mg/ml 5HT-BSA conjugate with 1:4,000, 1:10,000, 1:20,000, 1:40,000 and 1:80,000 dilutions of the rabbit anti-serotonin antibody overnight at room temperature with mild agitation. In parallel, equal volumes of these dilutions in which the 5HT-BSA conjugate is omitted are also incubated under the same conditions to provide positive controls. DAY 2 All samples are spun down at 14,000RPM for 15 minutes at 4oC Supernatant is collected for use in standard 5HT labeling protocol.