MAP Kinase Resource Forum

Hey! I just recently started phospho westerns. I was using PBST for the wash, as I usually use for yn other westerns, and my blots do not look good. Someone told me, that I should replace PBST with TBST for the phospho blots in order to improve results. What do you think about it? THX in advance!

Yest it is better to use TBST for western blots if you're looking at phosphorylated proteins. Depending on your antibody, the phosphate in PBSt could create problems, so it is always better to avoid it. Actually why not use TBST for all your blots? That way you don't have to make two buffers.

I would recommend the procedure here: http://www.mapkinases.eu/protocols/western-blot/

I am quite sure you will get your results, if you follow that!

Good luck!

thanks, it works much better now! well I actually changed an antibody too, but I am also doing it by your protocol!

I never use PBST, not for regular blots as well.