I'm running a C-elisa for Brucella in swine samples and even though from the eye point of view the controls seem ok, the OD (at 450nm) of wells designated as conjugate (cc) are not getting anywhere close to the indicated range by the manufacturer for the assay to be valid when calculating the percent inhibition. The protocol indicates the addition of sample dilution buffer in the cc wells.
i definitly agree with Emma Dale you have to calibrate your ELISA reader if not calibrated and be always stick with the method. I already used svanova cELISA kit and cc control never being out of range and that may be a problem in the reader so check it and i advice to verify it
Dear Sammy,
Try a correction at 540nm/570nm.The values given by the manufacturer may not always be accurate.
If you can do a scan across a range from 400 - 490 nm you might be able to find a better peak for you to use for the C-elisa.
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