Cytokine analyses?
Cytokine analyses?
Last Updated on Saturday, 19 June 2010 01:30 Written by Administrator Saturday, 19 June 2010 01:30
Question by sesame: Cytokine analyses?
I would like to know more about cytokines, more specifically how to quantify them. Is this the same as an immunoassay?
What is ELISPOT? And ELISA?
Please, if anyone could help with this, I’d be most grateful. I have attempted to research myself but I just don’t get it!
Best answer:
Answer by Mark S
ELISPOT and ELISA assays are both immunoassays that can be used to quantify cytokines and many other cellular products. The assays can be done in several ways, but one way is to put an antibody to the protein of interest (in your case, a cytokine) down onto a 96-well plastic plate–the plate is specially made to bind proteins. Then growth media from a cell culture solution is put into the wells. Any cytokine present bind to the antibodies. A second antibody to your protein is applied; it will bind to the cytokine also. Then a third antibody, which reacts with the second one but which also contains an enzyme that produces a colored product, is added. So in those wells that had cytokine-containing solutions you have a sandwich of first antibody–cytokine–second antibody—third antibody+enzyme. When the reactants for the enzyme are added, only those wells containing the sandwich will light up. The ELISPOT assay works similarly, except in this case rather than applying supernatants from a cell culture, cells are plated directly down onto the plastic (or sometimes nitrocellulose) surface and allowed to develop into colonies. Those colonies which are secreting cytokines will show up as spots (hence the name). I’ve provided links with more detailed information below.
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