Anti-EGFR antibody [ICR10] (ab231)

Hi,


I was wondering how I can find that whether the following anti EGFR products :







http://www.abcam.com/EGFR-antibody-EGFR1-Biotin-ab24293.html

http://www.abcam.com/EGFR-antibody-EGFR1-Biotin-ab24293.html



http://www.abcam.com/EGFR-antibody-F4-Biotin-ab24284.html

http://www.abcam.com/EGFR-antibody-F4-Biotin-ab24284.html




have inhibitory effect to block EGF binding or not. I would appreciate it if you could help me about it.

Kind Regards,

ANSWER:

Thank you for your enquiry and your interest in our products.

I can confirm that neither of these antibodies have been characterized for inhibition studies. ab24284 is suitable for IHC-P only; whilst ab24293 has been tested for Flow cytometry and IHC-Fr.

I have conducted a search for you and found another product in our catalogue ab231 which has been tested for inhibition studies. I would advise you to take a look at the on-line datasheet to see if it suits your need:

http://www.abcam.com/egfr-antibody-icr10-ab231.html

I hope this helps and if I can assist further, please do not hesitate to contact me.

Thank you for your email.

Can you please let me know, whether the product offered by you is the Final Mono Clonal Antibody (EGFR) or does it require further synthesis. As we are looking for Finish EGFR material for formulation, If it requires further synthesis, please advise us the company who can do it under sterile conditions.

Looking forward to hear from you soon.

ANSWER:

Thank you for your enquiry.

I am not sure I understand your question. Ab231 is a mouse monoclonal purified IgG prepared by ion exchange chromotography from ascites. Ab3103 is protein A purified, but I do not have the information on whether this was purified from ascites or from tissue culture supernatant. You should be able to use these as they are provided for any of the applications they have been tested in, which are listed on the online datasheets (I have included links below for your convenience).

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

As we discussed over the phone, I am sending information regarding the problem (not a puried lot of ICR10). Attached file (powerpoint) represents the data indicating the new batch of ICR10 (lot# 55227) is not purified or contamined with some species having absorption band around 280 nm. I also provide data showing the former ICR10 batch I used. I do not have the lot number for this because I finished the product and throw the vial. I used this ICR10 and got many live cell (HMEC cells) images of EGFR receptors. We use laser fluorescence microscopy and FRET imaging. Typically 20 nM of labeled antibody (direct staining) are used in our experiment and we have a set-up, which is capable of single molecule imaging (that is an ultra-sensitive fluorescence imaging technique). Please let me know if you further information. Thank you.

ANSWER:

I have just received some information from the originator of ab231 regarding the batch that you used. They re-tested that same batch and ran the antibody on both SPE and SDS gels. In both cases only one band was observed. They are confident regarding the purity of this product - the buffer is PBS with sodium azide added and there is no BSA or gelatin added.

As I mentioned in my previous email, our accounting department will be issuing you a refund for your order for this antibody. If I can be of any more assistance, please do contact me again.

Customer would like to know if these EGFR antibodies are specific for the truncated receptor.

ANSWER:

Thank you again for your patience. Regarding ab30, EGFR1 recognises the whole receptor protein rather than a truncated version. For ab5636, the synthetic peptide immunogen to produce the antibody is approximately 10 amino acid residues around EGFR [pY845]. The pY845 site is in the Tyrosine Kinase Domain of the Intracellular Domain. If the truncated EGFR protein includes the amino acid residues around 845 and the site is phosphorylated, the antibody will recognize the protein.

If you have any more questions, please do contact us again.

Customer would like to know if these EGFR antibodies are specific for the truncated receptor.

ANSWER:

Thank you for your patience. I have received the following information thus far:

Ab231: In Western blotting ab231 detects a 170kDa band which corresponds to EGFR. The epitope recognized by this antibody is located on the extracellular domain of the EGFR. I belive that the truncated form of EGFR lacks the cytoplasmic and transmembrane domain. If it is indeed this simple, then I would expect the antibody to recognise the truncated form. However, to our knowledge, this antibody has not been tested for reactivity with this form of EGFR and therefore we would be unable to offer any guarantees.

Ab2430: Ab2430 is made using a peptide corresponding to AA 1005-1016 of the human EGFR precursor. There are several truncated forms of EGFR and one of the more prevelant is EGFRvIII where amino acids 6-273 are replaced by a glycine residue. This antibody will recognize this variant, but it will also recognize the wild type. The researcher will need to compare the variants sequence to that of the wild type to make sure that this antibody's epitope has not been deleted.

Once I receive more information regarding ab5636 and ab30, I will contact you again.