Question (16449) | Anti-Cellubrevin antibody (ab5789)

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Question

we have purchased the rabbit polyclonal antibody to VAMP3 (Abcam ab5789) and used it for immunocytochemistry at the dilution 1:1000 and at our surprise we did not observed punctuate-vesicle structures only in the cytoplasm, but also a similar, but stronger, pattern in the nucleus. With the confocal microscope we also made Z-stacks, to exclude that there was only a surface labelling of the nucleus. We are questioning about the specificity of this antibody. I would be very greateful for your reply.

Answer

Thank you for your email. Ab5789 was characterized for use in Western blotting and was not tested in other applications. By Western blot, the antibody detects a band of approximately 13 kDa (predicted molecular weight: 12 kDa) representing recombinant rat VAMP-3. The immunogen used to generate this antibody was a synthetic peptide: STGVPSGSSAATGSNRRC, corresponding to amino acids 1-18 of Mouse VAMP3. Ab5789 has not been tested for cross-reactivity with the other VAMP isoforms. However, if you BLAST the peptide sequence used to generate this antibody (the ExPASy and NCBI links are on the online datasheet) there is no identity with other VAMPs. We do have a review from a customer who used this antibody in WB and IF (you can read her comments by clicking on the Abreviews tab located on the online product datasheet). She states that the antibody worked well for WB, but states "I expected the VAMP3 AB to have a more localized perinuclear staining, but the staining of this AB is homogeneous throughout the entire cell." It does seem that this antibody may not be suitable for IF. I do suggest that you submit an Abreview via the product datasheet. We always encourage customers to feed their results back to us, whether positive or negative, and make all information about each of our products available to researchers. If you send us an Abreview, we can offer you 50 Abcam Points (a further 100 Abpoints will be offered for an image). To find out more about our Loyalty Scheme and Abpoints, please visit our website and look at Your Account panel (left bottom section on our home page) or the Abreviews section on the product datasheet. You can submit the Abreview if you click on the Abreview section (top menu bar) on the on-line datasheet of this antibody. It is a very straightforward process. Should you need any help how to submit an Abreview, then please do not hesitate to contact us again. I hope this helps. Please contact us again if you have any additional questions.

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