NF kB mobilisation assay - help needed (Dec/13/2006 )
Am planning to set up this assay. first I'd like to replicate the expt in the paper, and then use my analog molecules to see if i can inhibit the mobilization.
Journal of immunology, 2001 167:366-374.
while the paper mentions the protocol a bit, I was wondering if I could get a dummy's guide to doing it, along with trouble shooting guidelines
I've not used this type of assay but it's basically immunohistochemistry on fixed and permeabilised cells. There's a similar in this newsletter from BD Europe (see link below) on page 24 with some methods on page 22-23. Obviously you don't need their machine just a (fluorescent) microscope and camera depending if you use fluorochrome-conjugated or HRP-conjugated secondary abs.