Problems with the nitroblue tetrazoleum (NBT) assay - (Oct/10/2005 )
Hi all,
I have performed the NBT assay to determine the effects of different compounds on the differentiation of leukemic cell lines. Although I've kept the final concentrations of NBT and PMA within published ranges (0.1% NBT, 0.1-1ug/ml PMA), I don't see anything close to what has already been published for my control samples. The NBT and PMA stocks were freshly made and are stored at -4C and -20C, respectively, so they should not be causing the problem. The incubation periods for the cells in the NBT/PMA solution varies from 15min to a few hours depending on the protocol, but prolonged incubations have not resolved the problem as even untreated cells begin to die. Has anyone else experiences this or know how to resolve this issue? Thanks.
OK, I figured it out. I changed the diluent for NBT and the cells responded as expected in the assay.
I have performed the NBT assay to determine the effects of different compounds on the differentiation of leukemic cell lines. Although I've kept the final concentrations of NBT and PMA within published ranges (0.1% NBT, 0.1-1ug/ml PMA), I don't see anything close to what has already been published for my control samples. The NBT and PMA stocks were freshly made and are stored at -4C and -20C, respectively, so they should not be causing the problem. The incubation periods for the cells in the NBT/PMA solution varies from 15min to a few hours depending on the protocol, but prolonged incubations have not resolved the problem as even untreated cells begin to die. Has anyone else experiences this or know how to resolve this issue? Thanks.
Hi I want to use NBT in an agar overlay method.
That is I want to overlay a TLC plate with agar mixed with cells (bacteria) to see if any of the TLC fractions can inhibit my test bacteria. Could you kindly give me an idea or reference