in gel kinase assayes?

[Glenn Jenkins]

In article <32500561.6DFD at genectr.hunter.cuny.edu>, Mauro
<Rabino at genectr.hunter.cuny.edu> wrote:

> Hi all 
> is there anybody that can help me to find reference, protocols and tips
> about testing kinase activity directly in the gel?
> For what kinase have been this methods used? Do I put the substrate
> directly in the SDS-gel before polimerization and than renature or I run
> a native gel?


Look up MAP kinase on medline since it is usually assayed in that manner.
Briefly, myelin basic protein is polymerized in the the gel and the
lysates are electrophoresed. Afterwards, the proteins are denatured
completely with GuanHCL and renatured ON in the coldroom followed by the
assay.

Hope this helps,
   Glenn Jenkins