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Position: Discussion forums > Bioscience biotechniques discussion forums > Immunology forum and cytometry forum
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ELISA weak reaction
Posted by: wageh (IP Hidden, New member, 1)
Date: April 17, 2005 08:57AM
Hi all
I am working on human blood sample to isolate leukocytes then challenge with lipopolysaccharide to produce cytokines namely TNF and IL-1 and finally measure the cytokine level by Sandwich ELISA. For months everything going well but recently I got many experiment results with weak ELISA reactions comparing with previous experiments I think I have a problem with ELISA not with the monocytes isolation technique because I got that weak reaction even with the standard cytokine that comes with the ELISA kit So could any one have a good experience with Sandwich ELISA comment on what in ELISA I have such a problem any comment will be much appreciated
Re: ELISA weak reaction
Posted by: nino (IP Hidden, Unregistered user, )
Date: June 23, 2005 08:45AM
hi all
I work in laboratory. I use Enzyme-Linked Immunosorbent assay Kits to measure FSH level in human blood.The principale of test is ,,two step,,sandvich-type immunoassay.On the last stage of an assay when the cromogen solution is added the wells are colourless. I need you advice what is the reason of it,I keep all condition which are done in FSH protocol.Please let me deal with your experience i need it very much sincerely yours Nino Bekaia
Re: ELISA weak reaction
Posted by: femmeauburn (IP Hidden, Advanced member, 115)
Date: June 23, 2005 11:22AM
Are the wells colorless just when the cromogen is added? Or do the wells remain colorless after incubation? It can take a little time for color to develop.
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