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RE: In Gel KAs



There are many kinases using GTP as well as ATP.
To determine the kinase you can try antibodies to blot or immunodeplete,
phosphatases to examine the regulation of the kinases, substrate specificity
using various peptide substrates. Good Luck
Yair Doza
__________________________________________________________
ASTRA CHARNWOOD
Discovery BioScience, Bakewell Road, Loughborough, Leics LE11 5RH, England
Tel: +44 (0)1509 645481   Fax: +44 (0)1509 645557
yair.doza@charnwood.gb.astra.com


> ----------
> From:
> Scott.Gridley@dartmouth.edu[SMTP:Scott.Gridley@dartmouth.edu]
> Sent: 	12 March 1999 10:16
> To: 	kinases@sdsc.edu
> Subject: 	In Gel KAs
> 
> I've got a partially purified kinase activity.  I know this kinase will
> utilize
> either ATP or GTP as its gamma p donor (is gtp usage uncommon?).  I also
> know
> that it will use myelin basic protein, casein, or histone H1 as a
> pseudo-substrate.  I have been attempting to get a molecular weight of
> this
> kinase through in gel kinase assays, but have been unsuccessful.  Does
> anyone
> have a good protocol, or any tips for optimizing in gels?  How about a
> good
> positive control?  I've tried casein kinase II alpha for that without
> luck...any info is much appreciated.
> 
> Scott Gridley
>