VIP (Vasoactive Intestinal Peptide) Antibody

We used VIP (product no. 20077) to stain for VIP positive neurons in the Pancreas and celiac mesenteric ganglion (CMG). For consistent and positive staining, we used a concentration of 1:500.

Animals were perfused with 4% PFA and tissue was post-fixed in 4%PFA overnight and then put into sucrose the following day. Tissue was sectioned at 20um on a Cryostat and mounted onto subbed slides.

Tissue was blocked with a 4% Goat Serum/Superblocker for 1hr. Primary antibody was added to the slides for 16hrs at room temperature. Slides were then washed 4 times for 15 min with a 1% triton X-100/Goat Serum/PBS solution. Secondary antibodies were then added for 3hrs at room temperature. Washed again 4 times for 15 min and then coverslipped.

Antibody labeled axon processes in CMG and cells in Pancreas.

Ty Redler, BS; OPS Sponsored Projects, Non-Clerical
Dr. Richard Johnson’s Immunohistochemistry Lab
University of Florida
College of Veterinary Medicine

We used the VIP antibody (Product ID: 20077) to stain for VIP positive neurons in mouse brain.

As recommended by Immunostar, after transcardial perfusion, brains were postfixed for 1.5 hours at 4°C in 4% paraformaldehyde. The following day, brains were sliced in the coronal plane on a vibratome at 40 μm and sections were collected in PBS. The tissue was incubated with primary antibody between 18–24 hours at 4°C, using 0.3% triton X-100, and 1% blocking serum.

We found that the most consistent staining for immunofluorescence was achieved with a 1:1000 dilution of the primary antibody. We evaluated antibody staining in the cortex and suprachiasmatic nuclei. The antibody strongly labeled cell bodies and also stained neuronal processes. Staining patterns matched previously published results.

Marina Silveira, Ph.D. and Michael Roberts, Ph.D.
Kresge Hearing Research Institute
University of Michigan

We have used this antibody to stain for VIP positive neurons (also stains processes) in frozen tissue sections and whole mount preparations of mouse intestine. We get consistent staining at 1:750.

Tissue prep:
4% PFA/PBS for 1.5 hrs, cryoprotected in 30% sucrose until they sink, OCT embedded, and sectioned.
For whole mounts, we open up intestine and fix in 4% PFA/PBS for 1.5 hrs.

IF protocol:
Sections:
Rehydrate in PBS, block with HINGS, stain in primary @4C O/N.
Wmts:
Wash in PBST, stain in primary with HINGS and DMSO @4C 48 hrs.

Sections:
Wash with PBS, stain with Alexafluor secondaries (Invitrogen), wash with PBS, and DAPI mount.
Wmts:
Wash in PBST, stain in secondary with HINGS and DMSO @RT 48 hrs.
Optional: proceed with MeOH dehydration and optical clearing.

Meenakshi Rao, MD PhD
Assistant Professor of Pediatrics
Columbia University Medical Center
New York, NY 10032