IUBio

Protein Labeling? Casein Kinase?

Donny Wong wong5 at fas2.fas.harvard.edu
Tue Jan 28 20:19:41 EST 1997


Hi everyone,

Does anyone know of a method of labeling proteins (radioactively or
without radiation) that doesnt involve iodination?  I'm worried about the
dangers of working with iodine (a gamma emitter) and I can't metabolically
label with 35-S Methionine because i'm giong to be working with proteins
that have already been purified.  

I found one possibility...casein kinase (I or II) which will phosphorylate
serine or threonine residues with 32P-ATP.  However, I don't know what
consensus sequence the enzyme recognizes (and hence, I may not be able to
use this method if my protein does not have the consensus site).  so does
anyone know of any other ways to non-specifically label proteins?

P.S.  I'm planning to do a far-western blot, in which you probe a membrane
onto which you have transferred protein with a labeled protein rather than
with antibody.  if anyone has any experience with this technique, i'd love
to hear from you.

thanks!!

donny
wong5 at fas.harvard.edu



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