How to find NFKb signalling is active in a cell line??? - (Oct/15/2010 )
could anybody suggest me how to find whether the NFKb signalling is active in a cell line or not???
Could you try transfecting in an NFKb reporter vector? http://www.promega.com/tbs/9pie849/9pie849.pdf
The above vector has an NFKb response element fused with firefly luciferase. The response element can be activated by stimulating your cells (eg with TNFa) and then you read the luciferase expression.
NFkB is a complex of various subunits and it is responsible for more than 200 gene expression.
Can you be more specific on what you are trying to find out about ?
easiest would be to see phosphorylated status of p65 subunit with ELISA based method.
As p65/p50 is reported to be most active and naturally occurring NFkB subunit