antibody works for wb not for elisa

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antibody works for wb not for elisa

Postby anosh123 » Apr 18 2015 10:02 am

Hello all!
I have a question regarding my primary antibody. This is a commercial antibody and I used it for both Western and ELISA. It works really well for Western but somehow did not work well with ELISA. Is this possible or it is some technical errors on my part?
anosh g
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Re: antibody works for wb not for elisa

Postby mdfenko » Apr 20 2015 11:40 am

look at the data sheet for the antibody. it will tell you what methods for which the antibody is certified.

the antibody may be to an epitope which is exposed by denaturation so won't work with a native technique like elisa.
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Re: antibody works for wb not for elisa

Postby r.rosati » May 13 2015 2:05 pm

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Re: antibody works for wb not for elisa

Postby rlw.altogen » Jul 08 2015 7:04 pm

Hi r.rosati, in case you were still wondering:

The WB uses denatured proteins, and so the epitope the primary antibody is design for may only be fully exposed after denaturation. As ELISA uses complex proteins the epitope might be obscured or hidden and thus the antibody won't work in the same way. Usually the manufacturer will specify if it works for either one or both assays.

However, if the antibody is designed for both but you're getting weak results for the ELISA I'd suggest going over your protocol. ELISAs require special attention to washing stages and incubation periods. If you need any help trouble shooting your ELISAs let me know, I do them a lot!
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Re: antibody works for wb not for elisa

Postby Jessica666 » Oct 15 2015 7:54 am

The instruction book ,generally, will note that which method it can be applied to .
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