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Description
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.