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Md. Toheder Rahaman 08-01-2013 04:12 AM

Problem in ELISA test
 
Hi all

I've got ELISA kit for Beta lactoglobulin (it is a milk protein). Company provided me pure beta lactoglobulin (standard), Coating Antibody and HRP conjugated detecting antibody. I have tried several times but I did not get good standard curve which is expected according to the protocol provided by the company.

Is there any one who can help me !!!

danfive 09-02-2013 08:24 PM

Re: Problem in ELISA test
 
Post the protocol.

Coating antibody to the plate is simple enough, but quality of the buffer and the accurate pH is important, so fresh buffer is key.

Similarly the buffer for diluting the standard is also important as well as the mixing, I suggest gentle pipette mixing at first, to see how the standard protein performs.

Lastly the blocking step is critical, it blocks any sticky sites on the plate surface (plastic binds protein naturally, so empty spots need to be coated with inactive protein).

Provide more details if you want specific help.


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