Anti-EpCAM antibody [Ber-EP4] (ab7504)

TAG72 antibody, ab16838, Lot GR99749-1: 66.6 ug/ml


Epcam antibody (Ber-EP4), ab7504, Lot GR103717-1: 47.3 ug/ml

Thank you for contacting us.

Unfortunately, this antibody has not been purified and therefore, its concentration has not been determined. However, we estimate that the antibody concentration in this tissue culture supernatant is 1-3 mg/ml.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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.

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

https://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.

https://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.

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

https://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.

Thank you for contacting us.

I was not able to find a purified, concentrated formulation of the Ber-EP4 clone in our catalogue, and we have not had it in the past.

Please do not hesitate to contact us if you need any more advice or information.
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Thank you for contacting us. As shown on the datasheet of the anti-EpCAM ab71916, www.abcam.com/ab71916, SW480 whole cell lysate is a recommended positive control. As explained on the phone, to our knowledge ab7504 has been tested in Western Blot. This monoclonal may work in this application and some optimisation may be needed. Since this antibody works in IHC, it may recognise an epitope made up of non-contiguous amino acids in their native conformation. Although the amino acids of the epitope are separated from one another in the primary sequence, they can be closer to each other in the three dimensional structure of the protein. If this hypothesis is correct, the antibody will only recognize the epitope as it exists on the surface of the folded structure. It is imperative in these circumstances to run a western blot in non-denaturing conditions, and this will be noted on the datasheet in the applications section. In general, a non-denaturing condition simply means leaving SDS out of the sample and migration buffers and not heating the samples. Certain antibodies only recognize protein in its non-reduced form i.e. in an oxidized form (particularly on cysteine residues) and the reducing agents β-mercaptoethanol and DTT must be left out of the loading buffer and migration buffer (non reducing conditions). We recommend to perform a western blot using a buffer without ionic detergents (e.g. SDS) and ideally without non-ionic detergents (e.g. Triton X-100). Cell lysis with detergent-free buffer can be achieved by mechanical shearing, often with a Dounce homogenizer or by passing cells through a syringe tip. In these cases a simple Tris buffer will suffice. Do not heat the samples. For more details I would suggest to look at the protocols book : www.abcam.com/protocolsbook or the Western Blot guide https://www.abcam.com/ps/pdf/protocols/WB-beginner.pdf . I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for contacting us in regards to Anti-EpCAM antibody [Ber-EP4] (ab7504) and Anti-EpCAM antibody [HEA125] (ab46714). As discussed over the phone yesterday, we do not currently have any information regarding these antibodies having been used in Western blotting but I have seen information relating to antibodies produced from these clones having been used for this application. It therefore seems likely that they should be able to be used for this application. If you would like we could take a look at the protocol you have been using to perform Western blotting in order to see if there are any suggestions we can make to improve the likelihood of seeing the desired band. I have attached a questionnaire to this email for this purpose. However, the Invitrogen Dynabeads you have purchased work like immunoprecipitation and it may be that the antibodies from these clones are more appropriate to bind to the native epitope of the protein, not the denatured form detected in the Western blot. I hope this information has been of help.

Thank you for contacting us. THE A431, HCT116 and MCF etc express this target so lysates of these can be used as positive control. For more information about the tissue that expresses this target please check Human Protein Atlas. As discussed; SW480 cell line can also be used as positive control. However we do not have tested results of this cell line which we can forward you for comparison. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for contacting us. As discussed on phone the antibody in question can be used with ab7504. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.