TBST vs PBST in western

TBST vs PBST in western

Postby kinoob » Sun Jul 17, 2011 8:30 pm

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!
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Re: TBST vs PBST in western

Postby cicenasj » Mon Jul 18, 2011 3:04 am

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!
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Re: TBST vs PBST in western

Postby kinoob » Tue Aug 16, 2011 1:37 pm

thanks, it works much better now! well I actually changed an antibody too, but I am also doing it by your protocol!
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Re: TBST vs PBST in western

Postby DNAer » Fri Feb 15, 2013 11:55 am

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