I did BrDU ELISA on gall bladder cell line to check effect of an infection ( treatment ). I found greater OD in infection than negative control ( only 2% serum ) but no differences in positive ( 10% FBS ) and negative controls. Rather in some cases negative wells showed higher ODs than positive control. After removing BrDU labelled media I neither washed the plate nor washed the plate before adding primary antibody according to Merck-Millipore kit protocol. Next time what should I do ensure validity of positive control or what other alternative positive controls I can use in BrDU ELISA ? ( I did not find mention of separate positive controls in most of the relevant papers).
Can higher FBS conc. affect BrDU incorporation into proliferating cells inversely or how ?
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