Anti-CARD10 antibody (ab36839)

Did you offer a sheep raised antibody to this product?

ANSWER:

Thank you for contacting us regarding this. I could not find any historical data showing that we had offered an anti-Card10 or anti-CARM3 raised in sheep. I hope this information is helpful. Please let me know if have any questions.

I am interested in your CARD10 antibody, as I have recently published a　paper on CARD10 (also known as CARMA3) knockout mice (See GENES &　DEVELOPMENT 21:984-996, 2007). However, upon reading the data sheet for the antibody, I discovered two significant problems with the antibody: the first is that EL4 cell lysate is considered to be a positive control, and the second is that the molecular weight appears to be　around 100 kDa.　First off, CARMA3 is expressed only in NON-hematopoetic cells, and　therefore would not be present in EL-4 cells, which are a mouse T cell　line (however CARD11/CARMA1 is expressed ONLY in hematopoetic cells, and　therefore it may cross-react with the antibody). Secondly, CARD10/CARMA3 (as well as CARD11/CARMA1) are much larger proteins than　100 kDa, they actually run between 130-150 kDa. Thus, I suspect that　the band indicated on the data sheet for the CARD10 antibody is not　correct, and wonder if you might be able to generate any other　antibodies that can detect endogenous CARD10. If so, I am certain that　my lab, and those of our competitors, would use the antibody a lot,　generating profit for your company. I hope that you can respond to this　email soon and let me know. Thank you very much.

ANSWER:

Thank you for your enquiry.

I have enquired at the originator of the product and they have confirmed that the immunogen for this antibody has no homology with CARD11 and a BLAST search does not yield proteins that large, so cross-reaction with linear epitopes of other proteins is highly unlikely.

We typically screen a panel of tissues and cell lines and deem any that are in the approximate size range based on the predicted MW (116kDa for CARD10) and the banding patterns given to be positive. The smaller size band seen on our data sheet could be a splicing variant, cleaved fragment, or some other modification. The markers we used are dyed and can run larger making the proteins appear smaller.

Unfortunately, we do not have plans to develop another antibody at this time. I hope the information above will be useful to you. Should you require any further information, please do not hesitate to contact me.