Removing phosphates - (Dec/10/2009 )

Hi, there!

I would like to know if anybody has tried to remove phosphate postranslational modifications in proteins. Till now I just knew lambda phosphatase method. Moreover, I'm trying to use lysis bufffer without phosphatase inhibitors (NaF and Vanadate, for example). I think I read sometime ago something at this respect about EDTA. Maybe somebody knows an special buffer which impairs phosphorylation... Does anybody have an idea?

Thanks very much!

EDTA is known to inhibit metalloproteases. May also chelate ions that are co-factors of certain enzymes (For example If you are gonna use CIP you should avoid EDTA in your buffer).

Maybe you can try to use just PBS and NP40 (0.1-1%) (without any phosphatase inhibitor NaF, Sodium orthovanadate, EDTA...)

Thanks, quiyo.

laurequillo on Dec 11 2009, 11:53 AM said: