lysis buffers, zymography and EDTA
ladasky at my-deja.com
Sat Nov 9 01:40:37 EST 2002
Jen <jrobertson1 at sympatico.ca> wrote in message news:<40gesu0jmd6o6gl3qr1ja16cp5ri4js25q at 4ax.com>...
> I use a standard PLC lysis buffer containing EDTA. I'm making whole
> cell lysates for zymography.
> (1) I'm assuming that the EDTA would kill the enzymatic activity of
> the proteins I'm interested in (bad thing)
> (2) can I re-make the lysis buffer and just omit EDTA?
There's another person in the newsgroups who asked a question about
lysis buffers today. I answered her and you may be interested in my
I use an EDTA-free lysis buffer in my work, and I provide the recipe.
What exactly is your "standard PLC lysis buffer"?
As far as I know, the main point of including EDTA in lysis buffer is
to inhibit divalent cation-dependent proteases. Other protease
inhibitors may be able to substitute for EDTA. Is your enzyme of
interest itself a protease? That could get complicated.
John J. Ladasky Jr., Ph.D.
Department of Biology
Johns Hopkins University
Baltimore MD 21218
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