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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?



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.




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