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Question about ELISA assay - (Apr/14/2015 )

I want to test for a particular antibody in a serum sample. I place the antibody in a well that has antibody specific antigen bound to the bottom. Then I add an enzyme labeled antibody that is specific to the serum antibody. This generates a colored solution, indicating the presence of the particular serum antibody. Am I doing it correctly?


you'll have to give more information about the procedure.


do you bind the antigen to a membrane?


how do you eliminate unbound antibody?


do you add a substrate to generate the colored solution?


The ELISA experimental principle

Utilizing labeling technology to mark antibodies(antigens) with enzyme and then make antibodies and antigens in the samples to be tested have a specific response with Enzyme-labeled antibodies (antigens). When meet with enzyme substrate, enzyme can catalyze and decompose substrate efficiently and exclusively and then it will produce colored products. According to the intensity of color, the specific antigens(antibodies) and quantum of antigens (antibodies) can be tested in the samples to be tested.

-Venus Timberlake-

Need more infoohmy.png


I second the notions. You should have wash and block steps in there to mitigate nonspecific reaction.

-Michael Starr-

how do you eliminate unbound antibody?unsure.png

You should have wash and block steps in there


-Zoe Winslet-