Anti-CETP antibody [ATM192] (ab2726)

One customer have bought one vial ab2726(CETP antibody ), the lot No is 72205. She used it to test CETP expressed in COS7 cells( transfected with full lenght human or animal CETP gene) at 1:400(recommendated by datasheet). But the result was not good: no bands with short exposure time, then elonged to 15mins, several bands appeared with high backgroud. the detect system is proved to work with other primary antibody. Then, she used another CETP polyclonal antibody prepared by themself to test as contrast. the result is good, clear target bands can be showed( see attchment for result picture) . Can you give me your suggestions to solve this problem asap?

ANSWER:

I'm sorry to hear your customer is having a problem with ab2726. Could you please tell me what blocking buffer was used (time and temperature too) and what primary incubation buffer was used too (with time and temperature). Has the customer tested the secondary (anti mouse antibody) with other antibodies and on its own?

Thank you for your patience, I look forward to hearing from you,

One customer have bought one vial ab2726(CETP antibody ), the lot No is 72205. She used it to test CETP expressed in COS7 cells( transfected with full lenght human or animal CETP gene) at 1:400(recommendated by datasheet). But the result was not good: no bands with short exposure time, then elonged to 15mins, several bands appeared with high backgroud. the detect system is proved to work with other primary antibody. Then, she used another CETP polyclonal antibody prepared by themself to test as contrast. the result is good, clear target bands can be showed( see attchment for result picture) . Can you give me your suggestions to solve this problem asap?

ANSWER:

I'm sorry to hear your customer is having a problem with ab2726. Could you please tell me what blocking buffer was used (time and temperature too) and what primary incubation buffer was used too (with time and temperature). Has the customer tested the secondary (anti mouse antibody) with other antibodies and on its own?

Thank you for your patience, I look forward to hearing from you,

Customer would like to know the following regarding ab2726: Is it a kappa or lambda? What is the affinity? Was it used in direct or coating ELISA?

ANSWER:

The antibody is IgG1. We have not further characterized the antibody in terms of Kappa and Lamda.

Here is a refernce for ELISA: A.P. Thomas Mouse Monoclonal Antipeptide Antibodies Specific for Cholesteryl Ester Transfer Protein (CETP). Hybridoma 15(5): 359-364, 1996.