Phosphorylation of Proteins - Detection?

Emile Schiltz eschiltz at sun2.ruf.uni-freiburg.de
Tue Jul 12 09:16:06 EST 1994

acer (forrest at biosci.uq.oz.au) wrote:
: G'day all,
: 	Does anyone know of a NON-radioactive procedure for testing whether a 
: protein is phosphorylated or not? We are interested in whether our recombinant
: is being phosphorylated when expressed in baculovirus. We've thought about using
: P32 in the culture medium but this is a messy and dangerous technique, does
: anyone know of a non-radioactive technique for determining whether our protein 
: is phosphorylated?

: Thanks for any replies,
: 			Alistair Forrest

: forrest at florey.biosci.uq.oz.au

hello Alistair,

there is a way: if you purify your protein and find somebody in the
neighborhood that can do electrospray-massspectrometry for you,
then you have a non-radioactive procedure, very sensitive also.
Other methods would be demanding more material than usually available
since you think of P32: diagonal electrophoresis or chromatography
with alkaline phosphatase inbetween.


Emile Schiltz <eschiltz at sun1.ruf.uni-freiburg.de>
           or <schiltz at bio2.chemie.uni-freiburg.de>
Institute of Organic Chemistry and Biochemistry
Albertstrasse 21, D-79104 Freiburg, Germany
phone: +49 761 203 6090

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