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?

Thank-you!

Hi tas,

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.

Cheers,

Scott

You could also run western blots for phospho-IKK and IKb. Demonstrated pathway activity would further flesh out the luciferase experiments mentioned above.