Anti-Vimentin antibody [Vim 3B4] (ab28028)

Inquiry: I'm looking into a panel of antibodies from your company to use in cells of pig origin. ab758 ab27988 ab7504 ab124687 ab28028 ab11333 ab32362 I would like to know the following: Have they been tested in pig cells? If yes, are they reactive towards pig cells? If you do not have that information, can you tell me the epitope or peptide sequence that was used in generating that antibody? If you do not want to tell me that info, can you tell me the identity percentage of match of the pig compared to the epitope that it was used in generating the antibody. Many thanks! S

ANSWER:

Thank you for your inquiry.

For ab758, the immunogen that was used was cytoskeletal extract of human hepatocellular carcinoma cell line, Hep3B. We have not validated this antibody in pig. But the human sequence is 85% homologous with the pig cytokeratin 8 sequence. We generally say anything with >85% homology would be expected to cross-react, so this is right on the edge. However, we do have two anti-cytokeratin 8 antibodies that we know do work in pig as ab2531 and ab2530.

http://www.abcam.com/Cytokeratin-8-antibody-C-51-ab2531.html

http://www.abcam.com/Cytokeratin-8-antibody-C-43-ab2530.html

For ab27988, this antibody has been validated in pig and we therefore guarantee it will work for pig in IHC-P, IHC-Fr in accordance with our Abpromise.

For ab7504, the immunogen was human breast carcinoma cell line, MCF-7. Pig and human EpCAM is 83% homologous, so this is a little low. We have an anti-EpCAM that is predicted (>90% homology) to cross-react with pig, and that is ab71916. However, this hasn't been validated and therefore not guaranteed in pig.

http://www.abcam.com/EpCAM-antibody-ab71916.html

I have to check with the lab on ab32362 and ab124687 since I don't have the immunogens readily available to BLAST.

For ab28028, full length native protein (purified): Vimentin isolated from bovine eye lens was used as an immunogen. Pig vimentin and bovine vimentin are 97% homologous, so there is a good chance this will cross-react with pig. However this hasn't been tested and therefore isn't validated in pig. We do have a few vimentin antibodies that work in pig though,like ab20346 and ab8069.

http://www.abcam.com/Vimentin-antibody-VI-10-ab20346.html

http://www.abcam.com/Vimentin-antibody-V9-ab8069.html

For ab11333, cultured human thymic fibroblasts was used as an immunogen. We don't have a particular protein sequence to look up for this one, so unfortunately we don't know how/if it will work with pig.

I hope this information helps. I'll be in touch again once I hear about ab32362 and ab124687.

Hello,
can you recommend me some anti-epithelial membrane antigen primary antibody, which can be used for immunofluorescence, western blot and immunohistochemistry from paraffin embedded section?
I need to characterized primary ovarian cells for epithelial or mesenchymal origin and want to do double staining with anti-vimentin primary antibody (I think about mouse ab28028 or rabbit ab16700).
Thank you
best regards

ANSWER:

Thank you for contacting us and your interest in our products.


We have several antibodies directed against epithelial membrane antigen (also known as MUC1), however only one has been teated in all the applications you wish to use it for. This antibody(ab28081) is a mouse monoclonal and would therefore be most appropriate to use for double staining with the rabbitanti-vimentin antibody (ab16700).


If you wish to use the mouse monoclonal anti-vimentin antibody (ab28028) then I would suggest usingthe rabbit monoclonal anti-MUC1 antibody (ab109185) which whilst it has not been tested with IF,immunocytochemistry has been performed using this antibody.


If you require secondary antibodies to perform this staining we have several to choose from which can easily be selected for using our secondary antibody search:


http://www.abcam.com/index.html?pageconfig=productmap&cl=918


I hope this information has been of help. If you require any further information please do not hesitate to contact us again.