Dear Charles,
A few years ago I used Wako Lys-C for digestion of a glycoprotein a
Leader derived from the Barrier Protease (BSD) of Saccharomyces
cerevisiae under reducing conditions and it worked very well. I used
0.2 amidase unit of Lys-C per 1.0 mg of BSD glycoprotein (approx.
40000 g/mol) dissolved in 0.9 ml of 50 mM Tris-HCl and 6.5 mM DTT, pH
8.9. The sample was left at 4C for 92-144 h, and then the digestion
was stopped with 50 ul of glacial acetic acid/ml sample.
Ref: Jars, M. U et al. J. Biol. Chem. Vol. 270, pp. 24810-24817, 1995.
Mette
______________________________ Reply Separator _________________________________
Subject: Re: Lys-C digest
Author: Ken Mitchelhill <k.mitchelhill@medicine.unimelb.edu.au> at Internet
Date: 06-02-98 09:25
Charles,
I tried this once with Wako Lys-C in 10mM DTT with a protein that is
usually digested brilliantly and it was a disaster.
Ken
>Does anyone know how well Lys-C digests work under reducing conditions?
>
>Charles McNemar, Ph.D.
>Associate Principal Scientist
>Schering-Plough Research Institute
>2015 Galloping Hill Road
>Kenilworth, NJ 07033
>
>908-298-7194
>FAX: 908-298-4844
>
>E-mail: charles.mcnemar@spcorp.com
********************************
Ken I. Mitchelhill
The John Holt Protein Structure Laboratory
St. Vincent's Institute of Medical Research
41 Victoria Parade
Fitzroy 3065 Victoria
AUSTRALIA
Telephone: 61-3-9288 2480
Facsimile: 61-3-9416 2676
Email: k.mitchelhill@medicine.unimelb.edu.au
Laboratory: http://www.medstv.unimelb.edu.au/WWWDOCS/SVIMRdocs/JHPSL.html
ABRF: http://www.medstv.unimelb.edu.au/abrf.html
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From: Ken Mitchelhill <k.mitchelhill@medicine.unimelb.edu.au>
Subject: Re: Lys-C digest
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