ELISA Test

About microscopic forms of life, including Bacteria, Archea, protozoans, algae and fungi. Topics relating to viruses, viroids and prions also belong here.

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Elersong
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ELISA Test

Post by Elersong » Wed Oct 29, 2008 7:29 pm

Does anyone know exactly how the antibodies/antigen molecules are bound to the inside of the microtitre wells in the first step of the ELISA? I have done some research and can't seem to find the mechanism by which they are attatched.
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chen.lie
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Re: ELISA Test

Post by chen.lie » Wed Oct 29, 2008 11:23 pm

Hello,

For the Indirect ELISA method, you can put x µL of your sonicated bacteria in the wells and let dry overnight at 37°C. Then wash the plates the following day before continuing the experiment.

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Post by blcr11 » Thu Oct 30, 2008 1:47 pm

I think it is mostly non-specific, electrostatic interactions. Many proteins spontaneously stick to plastics like polystyrene, less so to polypropylene. This is how the indirect ELISA plate is set up, as chen.lie described. Once you've bound the reagent of interest, you usually then have to block as much of any remaining potential non-specific sites by washing, then adding an excess of some general protein like albumin or gelatin, and then washing again.

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Post by Elersong » Sun Nov 02, 2008 11:32 pm

That is something that I didn't know. So there would be no heat fixation?

I guess a way to further the question is: What is it about the proteins that cause them to become attracted to the inside of the well?
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