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Question (40663) | Anti-Sonic Hedgehog antibody (ab19897)

Go to datasheet (ab19897)

Question

No signal response in sELISA, usinghuman recombinant protein as antigen.

Abcam community

Verified customer

Asked on Mar 20 2012

Answer

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.

Abcam Scientific Support

Answered on Mar 20 2012

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