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Re: detection




In my hands the P-Ser and P-Thr antibodies have never worked well, and I 
have seen very few publications using them successfully. In many cases, 
you can detect phosphorylation through the mobility shift of the 
substrate protein in SDS-PAGE. Alternatively, you can run the substrate 
on an isoelectric focusing gel or columm, because the incorporation of 
the phosphate causes quite a huge change in the IP.

Hope this helps,

Gerard

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Dr.Gerard Drewes                          Phone: (+49)(0)40-89984766 office
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On Mon, 15 Mar 1999, Vladimir Krystof wrote:

> Hello all,
> 
> can anybody tell me, if there is another detection possibility of 
> kinase activity except radiolabelled ATP?
> I think about antiphosphoSer/Thr antibodies, but don't  know 
> whether the immunodetection is sensitive and specific enough if we 
> use purified enzyme and add protein as a substrate to 1mg/ml.
> Thanks,
> 
> 
> 
> 
> Vladimir Krystof
> ---------------------------------------------------
> Laboratory of Growth Regulators
> Palacky University
> Slechtitelu 11
> CZ-78371 Olomouc
> Czech Republic
> 
> phone: ++420 68 5222487
> fax:   ++420 68 5221357
> 
> http://prfholnt.upol.cz:81/growthreg
>