I recently did WB to check the pCREB/CREB after glucagon stimulation in mouse liver, but I got no band both for pCREB and CREB. I used RIPA buffer with proteinase and phosphorylase inhibitor cocktail to do the liver protein extraction. Mix the liver lysates wtih Lameli buffer and BME, and run with reducing SDS-PAGE. Does any one ever have the experience to do the WB for pCREB and CREB in mouse liver? Do I need to do the nuclear extraction?

I am not sure if the problem is the specifity of the antibodies or the sample preparation.

Both pCREB (9198) and CREB (9197) antibodies are from CST.