NPY Antibody – Rabbit

Immunohistochemistry: 1:5,000-1:10,000 (biotin-streptavidin/HRP).

Dilutions listed as a recommendation. Optimal dilution should be determined by investigator.

Tissue Preparation:
10 um cryostats or 50 um vibratome
-Fixative: 4% paraformaldehyde in 0.1M phosphate buffer, pH 7.4; 500 mL over ~ 20 min.
-Post Fixation: 1.5 hour at 4°C in 4% paraformaldehyde in 0.1 M phosphate buffer, pH 7.4.
Note: If needed, low levels of glutaraldehyde (0.1-0.3%) may be used in conjunction with paraformaldehyde.

Immunohistochemistry/Immunofluorescence:
In rat central nervous system, the antiserum has significant staining with a very low background at a 1/5,000 – 1/10,000 dilution using the Biotin-Streptavidin/HRP detection method. All staining is blocked by pre-absorption of the diluted antiserum with excess NPY. Cross reactivity experiments in which diluted NPY antiserum was absorbed with excess peptide YY, avianpancreatic polypeptide, B-endorphin, vasoactive intestinal peptide, cholecystokinin or somatostatin showed no affect in blocking the intensity of staining. Incubate 18-24 hr at 2-8° C.