Measuring NFkB activation - (Jun/26/2005 )
Can anyone help me?
What methods are there for measuring NFkB activation other than EMSA and Immunofluorescence etc? Does anyone else have any other suggestions?
Have you thought about a luciferase assay. You make/buy/get a construct containing the NFkB responsive element upstream of a luciferase gene (works with other marker genes). Transfect it into cells and stimulate with drug/compound of choice and measure luciferase activation using a luminometer.
You could also run western blots for phospho-IKK and IKb. Demonstrated pathway activity would further flesh out the luciferase experiments mentioned above.