The antibody has a proven strong Biotin-Streptavidin/HRP staining at a 1/4000-1/6000 dilution in rat amygdala, cortex, and suprachiasmatic nucleus.
The specificity of the antiserum was examined by soluble pre-adsorption with the peptides in question at a final concentration of 10-5 M. VIP immunolabeling was completely abolished by pre-adsorption with VIP. Pre-adsorption with the following peptides resulted in no reduction of immunostaining: Secretin, gastric inhibitory polypeptide, somatostatin, glucagon, insulin, ACTH, gastrin 34, FMRF-amide, rat GHRF, human GHRF, peptide histidine isoleucine 27, rat pancreatic polypeptide, motilin, peptide YY, substance P, neuropeptide Y, and CGRP.
The histochemical antibody for VIP is generated in a rabbit against porcine VIP conjugated to bovine thyroglobulin with carbodiimide. The antibody is provided as 100 µL of lyophilized whole serum, and 0.09% sodium azide.
Photo Description: IHC image of rat cortex (above) and low magnification image of rat striatum and amygdala staining for VIP (below). The tissue was fixed with 4% formaldehyde in phosphate buffer, before being removed and prepared for vibratome sectioning. Floating sections were incubated at RT in 10% goat serum in PBS, before standard IHC procedure. Primary antibody was incubated at 1:5000 for 48 hours, goat anti-rabbit secondary was subsequently added for 1 hour after washing with PBS. Light microscopy staining was achieved with standard biotin-streptavidin/HRP procedure and DAB chromogen.
Quantity / Volume: 100 µL
State: Lyophilized Whole Serum
Reacts With: Avian (Bird), Bat, Bubalus Bubalis (Buffalo), Bullfrog, Canine, Cat, Chick, Chicken, Dog, Dove, Fish, Fowl, Fox, Frog, Gilt (Pig), Goldfish, Guinea Pig, Hamster, Hedgehog, Hen, Human, Lizard, Mink, Mole, Mollusk, Monkey, Mouse, Mudpuppy (Necturus Maculosus), Ovine, Pig, Pigeon, Platypus, Possum, Rabbit, Raccoon, Ram, Rat, Scorpaena Porcus (Fish), Sea Lamprey, Sheep, Shrew, Snake, Sparrow, Sparus Aurata (Fish), Squirrel, Starfish, Sting Ray, Suncus Murinus, Tadpole, Turkey, Turtle, Worm (Spionidae), Zebra Finch, Zebrafish
Availability: In Stock
Preabsorption Control: Available at Sigma Aldrich, Catalog # V6130
Alternate Names: Vasoactive intestinal polypeptide; VIP peptides, anti-VIP
Gene Symbol / ID, Accession #: VIP,7432
03/19/2015 - 01:48:03 PM
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.
Rehydrate in PBS, block with HINGS, stain in primary @4C O/N.
Wash in PBST, stain in primary with HINGS and DMSO @4C 48 hrs.
Wash with PBS, stain with Alexafluor secondaries (Invitrogen), wash with PBS, and DAPI mount.
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
08/22/2012 - 02:45:39 PM
Worked like a charm
It turned out that this antibody worked like a charm on my frozen sections. This is definitely an excellent VIP antibody.
Dr. David Ma, Research scientist
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Q & A Wall
If I take out 10 µL of VIP Antibody from the vial after it is reconstituted, how much do I need to add to dilute it out to 1/10?
Question By Jeff
As a best practice, we recommend customers reconstitute the newly opened vial with 100 µL of distilled or deionized water, and then add 900 µL of PBS or Tris buffer. The antibody will then be diluted to a 1/10 ratio. Then make aliquots based on intended use and freeze. One suggestion is to aliquot 10 vials with 100 µL each into small eppendorf or cryovials and store the vials you don’t plan to use soon (within 1 -2 weeks) in the freezer.
Therefore in your case, Jeff, if you remove 10 µL of antibody from the vial, you are left with 90 µL of antibody. To get the same 1/10 ration you need to add 810 µL of buffer to the 90 µL of antibody for a total of 900 µL. This would provide the same 1/10 dilution, or 90 µL antibody to the total of 900 µL both antibody and buffer.
Answer By Colleen on 2015-10-29 14:13:58
I am interested in your product 20077. My VIP protein has sequence: KAMLPRNGSQLLLLIALCSVLYTRTLSLPYPSM RRPTRHADGLFTSGYSKLLGQLSARRYLESLIGKRVSNELM EEQMPVKRHSDAIFTDNYSRFRKQMAVKKYLNSVLTGKRSQEDPSTLKEDSPRNDPPFPESYDDVSVDELLSHLPLV Please, can you check the epitope of your anti-VIP antibodies against my sequence for possible crossreactivity,
Question By Svetlana
VIP has been used effectively in buffalo, chicken, guinea pig, hamster, human, monkey, mouse, pig, rat, sparrow, sting ray, and zebrafish. Statistically it should work with your protein but as you know that is not always possible to predict. It is advisable to try a dilution series to determine the optimal antibody dilution to use. For example, if a product data sheet suggests using a 1:500 dilution, you might make dilutions of 1:50, 1:100, 1:500, 1:1,000 and 1:10,000, to determine the optimal dilution.
Answer By Technical Support on 2013-08-09 15:16:35
This product contains the preservative sodium azide. The concentration percent of the sodium azide is ≤ .09%. Although this hazardous substance is a concentration below that required for the preparation of a Material Safety Data Sheet, we created a standard MSDS for your records.Download MSDS