The Substance P antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat substantia nigra and spinal cord using biotin/avidin-HRP techniques. Recommended primary dilutions are 1/500-1/1000 in PBS/0.3% Triton X-100 - Cy3 Fluorchrome and 1/6000-1/8000 in PBS/0.3% Triton X-100 - biotin/avidin-HRP Technique. The specificity of the antiserum for Substance P was demonstrated using soluble pre-adsorption with the peptides in question at a final concentration of 10 μg of peptide per mL of diluted antiserum. Substance P immunolabeling was completely abolished by pre-adsorption with Substance P. Pre-adsorption with the following peptides resulted in no reduction of immunostaining: neurokinin A, neurokinin B, somatostatin and neuropeptide K.
