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Problem in ELISA test

Problem in ELISA test - Biology Forum

Problem in ELISA test - Biology Forums. Ask questions and discuss the study of Biology. If you have Biology questions from your homework ask them here!


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Old 08-01-2013, 04:12 AM
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Default 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 !!!
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Old 09-02-2013, 08:24 PM
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Default 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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