Neuropeptide Y Antibody

We have used your NPY(Product ID: 22940) antibody and love the results we obtain with this antibody. While it is not as clear as your Vasopressin antibody (Product ID: 20069), it works very well for our research purposes. For NPY we use a dilution of 1:3000 and amplify with tyramide solution at a 1:50 dilution. It is mixed with 10% serum, 10% biotin diluted in 80% PBS-T and incubated for 48-72 hours at 4 degrees C. We used brain tissue from Thamnophis sirtalis (Red-sided garter snake).

Above IFC image entitled "C." is NPY interneurons (RED) in mouse striatum, GREEN staining is GFP expressed by medium spiny neurons. NPY antibody was used at 1:1000 dilution with the staining protocol described below. Antibody performed very well. Scale bar = 25 microns. Immunohistochemistry: Mice were euthanized and perfused with PFA (4% in PBS), post fixed for 24 hrs, and their brains cryoprotected in 15% sucrose (24 hrs) followed by 30% sucrose (48 hrs). Coronal sections that included the corticostriatal, globus pallidus, or substantia nigra regions (30 um) were prepared using a microtome and then stored in PBS at 4C until processing. Sections were processed/stained in parallel as follows: Free floating sections were rinsed 3x with PBS and incubated 90 min at room temperature (RT) in PBS supplemented with donkey serum (5%) and Triton X-100 (1%). Primary antibody combinations were prepared as a master stock in PBS supplemented with donkey serum (2.5%) and Triton X-100 (0.5%) and applied to sections for 72 hrs at 4°C with gentle agitation. Sections were then rinsed 8x with PBS supplemented with Tween-20 (0.05%, at RT) and incubated with a secondary antibodies diluted in PBS supplemented with donkey serum (2.5%) and Triton X-100 (0.5%) for 1 hr at RT with gentle agitation, followed by 7 rinses with PBS and one rinse with deionized water. Sections were mounted onto slides and allowed to dry for ~18 hrs, after which cover slips were mounted with Vectashield and sealed with nail polish.