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The neuronal nitric oxide synthase C-terminal antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat hypothalamus, striatum, cortex and spinal cord using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/1,000 - 1/1,500 in PBS/0.3% Triton X-100 - Cy3 Technique and 1/8,000 - 1/12,000 in PBS/0.3% Triton X-100 - Bn/Av-HRP Technique. By Western blot analysis of brain homogenates the antibody specifically labels a band of approximately 155 kD. Immuno-labeling is completely abolished by pre-adsorption with synthetic human nNOS (1419-1433) at 5 µg per mL of diluted antibody. No cross reactivity with other forms of NOS were observed. The nNOS antiserum has been used successfully in human, rat, mouse, guinea pig, cat, and monkey tissue. Detection of nNOS from other species will depend upon sequence homology.