Anti-ID3 antibody (ab55269)

Thank you very much and have a great evening!

ANSWER:

I'm happy to help. Please contact us again if you have any further questions.

Hi,    Thank you so much! I will try it. You mean you test the ID3 antibody in tissue samples? did you test this antibody in cell lines? would you be so kind to tell me which cell lines have the higher expression of ID3? and I will be appreciated if you can give me some publications related to your ID3 antibody which they detected ID3 expression In cell lines?

ANSWER:

All the testing data we have using ab55269 can be found on the online datasheet for ab55269 on our website. The tissues I noted as expressing high levels of ID3 was information I found in SwissProt database of proteins. We have links to this and other external protein and genomic databases for each primary antibody on our website. Here is the link on our website that I used: http://www.uniprot.org/uniprot/Q02535 The information can be found under the 'General annotation (Comments) section. So we have not actually tested these samples with ab55269. We also note any publications using the product on its datasheet in the 'References' section. In this case we do not know of any publications using ab55269. I hope this helps. Please let me know if you have any further questions.

Thank you for your reply, Actually I use this antibody to do the Western blotting, the dilution is 1ug/ml in 5% BSA-TBST. and using regular RIPA buffer to extract the protein, loading 40ug protein per lane. Thank you!    By the way, if you know some publications showed that this protein expressed in other cell lines, please forward to me, so I can check with this antibody!

ANSWER:

I have found that ID3 is expressed abundantly in lung, kidney and adrenal gland, but not in adult brain.

Also, to increase sensitivity of detection, I would recommend using a higher concentration of antibody, such as 2 or 4ug/ml. Also, if you are not already, I suggest incubating the primary overnight at 4 deg C as well.

I look forward to hearing if these suggestions prove useful.

I bought an ID3 antibody from you company (ab55269),  I found in your website that this protein is expressed in Hela cells, but I tried several times, it can recognize the transfected ID3 protein in Hela cells, but can not detect any endogenous expression of ID3 in Hela cells. So, would you be so kind to tell me did you company try other cell lines to detect this protein with your antibody? or do you have any other clues that ID3 is expressed in Hela cells?    Because I want to detect the endogenous ID3 expression, and I tried several cell lines with your company's antibody and failed, so I want to whether it is because the antibody or specific cell lines? Thank you and have a great day!

ANSWER:

Thank you for your enquiry.

I think it would be helpful to get a bit more information on your protocol and sample preparation methods. For example, are you doing western blotting or ICC? What dilution of the primary antibody are you using? What methods of cell fixation and/or cell extraction are you using? If WB, how much protein are you loading on the gel?

Thanks in advance for the additional information.