Too high Tris-HCl in stacker SDS-PAGE a problem?
Ian A. York
iayork at panix.com
Sun Apr 5 14:39:50 EST 1998
In article <Pine.SGI.3.96.980405152136.17989F-100000 at umbc7.umbc.edu>,
zaxxon <zhowar1 at umbc.edu> wrote:
>Okay - here's my next question. Upon reviewing all of my solution-making
>notes, I found that I've been making up the stock solution for my stacker
>gel (for SDS-PAGE) with a higher than intended Tris-Cl concentration.
I haven't done the experiment, but my gut feeling is that the difference
you mention will not cause problems.
As a related point, anyone using the recipes in Harlowe's Antibodies: A
Laboratory Manual should check the numbers; there are several typos in
them. I can't remember the details, but the recipe for SDS-PAGE running
buffer may have been one of the ones that's wrong, and at least one of the
acrylamide gel concentrations is wrong.
I used wrong recipes a couple of times (and not to point the finger at
Harlowe, some of the mistakes were my own) before spotting the mistakes
and re-checking all the numbers, and still got adequate, if unspectacular,
gels. The system is robust enough to tolerate some leeway.
Ian York (iayork at panix.com) <http://www.panix.com/~iayork/>
"-but as he was a York, I am rather inclined to suppose him a
very respectable Man." -Jane Austen, The History of England
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