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Position: Discussion forums > Bioscience biotechniques discussion forums > Pharmacology and toxicology forum
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sandwich elisa trouble
Posted by: vdnadung (IP Hidden, New member, 1)
Date: November 9, 2007 06:25AM
Hi everyone,
I have trouble with sandwich elisa and i need your advice. I'm trying to establish a sandwich elisa kit to detect a bacterium. There is no problem with coating antibody (i test it by using indirect elisa) and with capture antibody (conjugated with HRP) (i test it by using direct elisa, coating the plate with purified antigen (a protein of this bacterium, not cell)). I used to test them by using sandwich elisa (the target antigen is that protein) and it's ok too. However, when i do sandwich with the cell (10^5, 10^6, 10^7 cells/well), the signal is very low (less than 0.6) although i increased the quantity of the coating and capture antibody up to 1microgam/well. I hope you give me some advice on this trouble. Thank you and best regards.
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