prtoteinase digestion

.Krimer Alejandro a.krimer at biosidus.com.ar
Tue Jun 15 11:40:25 EST 1999


	Dear Bernarºd:

	Trypsin is a calcium-dependent protease, you can stop the reaction
by adding EDTA 10 mM (final concentration) or some inhibitors (STI). You can
also use PMSF.

	In general the ratio protein:trypsin is ranging from 100:1 to 20:1
(w/w). The incubation time depens on the conditions, for example at 4 hours
at 37 ºC, 12-24 hs. at 25 ºC.

	Hope this helps,

	Alejandro Krimer 

> In article <19990610155335.68176.qmail at hotmail.com>, ips98 at HOTMAIL.COM
> (inderpal singh) wrote:
> 
> >       Does anyone know how to digest proteinase with trypsin and how  to
> 
> > terminate the reaction! If so,please write in detail, how much conc.  of
> 
> > protein and trypsin etc should be taken and for how long the reaction
> should 
> > be performed.
> 
> What protocols have you tried so far?  How did they work?
> What literature references are you following?
> 
> -- 
> Bernard P. Murray, PhD
> Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA



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