Anti-VIP antibody (ab6184)

Dear Abcam TEAM,

WeE do look for antibodies to build up an ELISA for human and rat VIP.

Do you have any infromation about this application.

What other antibodies would you recommend?

Thank you for your help Best Regards

ANSWER:

Thank you for your enquiry. To our knowledge this antibody has yet to be used in this application. All the information we have available is that which is listed on the product datasheets. We have no further information about this application. Best of luck with your research.

BATCH NUMBER 40881 ORDER NUMBER -- NOT SPECIFIED --

DESCRIPTION OF THE PROBLEM there are only few immunoreactivities and a very high grade of background staining

SAMPLE bovine (cow) abomasal wall

PRIMARY ANTIBODY dilutions: 1:500; 1:750; 1:1000 buffer for antibody dilutions: PBS / 0,03% TRX / 1,5% BSA

SECONDARY ANTIBODY rabbit anti-sheep 1:300;

DETECTION METHOD detection: ABC / AEC

POSITIVE AND NEGATIVE CONTROLS USED omitting the primary antibody detected no staining; positive controls with duodenum from rats showed specific staining of the plexus myentericus, but showed background, too.

SAMPLE PREPARATION formalin-fixed paraffin-embedded tissue; 6?m sections

TRANSFER AND BLOCKING CONDITIONS 20 min cooking with citrate buffer; 30 min H2O2 in methanol; 30 min blocking with 5% rabbit normalserum

HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes

ANSWER:

I'm sorry to hear you are having problems with ab6184. I would like to recommend the following: -an enzymatic antigen retrieval step -fixation using Zamboni's fixative (4%PFA + 15% picric acid).

Please let me know if these modifications to your protocol help your staining and do not hesitate to contact us if you need further advice,

I would like to confirm what you mean by "Specificity has not been examined, but results from immunohistochemistry indicate reactivity is similar to findings using other VIP antisera" which is written on this datasheet. Can you be more specific about what these other findings are please? Also can you please confirm that the antibody cross react with the species on the datasheet i.e. Cross-reacts with Human, Mouse, Rat, Cow, Dog, Guinea pig, Pig and Rabbit.

Thank you.

ANSWER:

Thank you fo your patience. We now have an answer to your initial enquiry.We can confirm that the antisera is suitable for use in all species identified on the datasheet, ie, Human, Mouse, Rat, Cow, Dog, Guinea Pig, Pig and Rabbit. The statement "Specificity has not been examined, but results from immunohistochemistry indicate reactivity is similar to findings using other VIP antisera" has been included on the datasheet to indicate that reactivity against other peptides has not been examined. For example, some VIP antisera appear to react with PACAP. However, our antiserum is a high-quality reagent that has been used extensively by many groups and provides results consistent with the accepted distribution of VIP throughout the nervous system, particularly sympathetic and sensory neurons. I hope this helps you with your research.

Does this antibody cross-react with chick VIP?

ANSWER:

We have not tested this antibody in chick.

Could you tell me if the antibody works in formalin-fixed and paraffin-embedded tissues or just with Zamboni´s fixative?

Could you also give me any reference that use this antibody in rabbits?

ANSWER:

As far as we are aware, the antibody has only been tested using Zamboni's, so we do not know if it will work with paraffin embedded tissues. The work with rabbit was carried out by the antibody's originators, a research lab in Australia, however it has not been published so we don't have a reference for this.