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No signal response in sELISA, usinghuman recombinant protein as antigen.
Asked on Mar 20 2012
Thank you for your call last week and for your patience while I have been in touch with my colleagues about these antibodies. I apologize for the delay getting back to you.
The capture antibody ab87382 was tested at a coating concentration of 1 ug/mL, which corresponds to a 1:200 dilution as the antibody is provided at 0.2 mg/mL. If the antibody was used at 1:1000, this is a 0.2 ug/mL dilution which might have contributed to a weak signal in the ELISA. I would recommend decreasing the concentration of this antibody to 1:200. The detection antibody ab19897 was tested at 0.5 ug/mL, which is approximately the same concentration that you have used (the antibody is provided at 0.6 mg/mL for the lot that you have).
We had also discussed testing the secondary antibody to see if it cross-reacts with the mouse capture antibody ab87382. Have you had a chance to test this?
We do guarantee these antibodies to work in your assay, so if you think that they are faulty, I would be happy to send a replacement or issue a credit or refund if you prefer. Please let me know if you have any questions, and keep me updated about any progress with these antibodies. Feel free to call 888-772-2226 and ask to speak to me anytime.
Answered on Mar 20 2012