can i use EDTA instead of TEMED for SDS PAGE ?

[Pow Joshi]

2008/5/29 DK <dk from no.email.thankstospam.net>:

> In article <mailman.251.1212073353.3533.methods from net.bio.net>, "Pow Joshi"
> <pow.joshi from gmail.com> wrote:
> >2008/5/28 DK <dk from no.email.thankstospam.net>:
> >
> >> In article <mailman.243.1212019971.3533.methods from net.bio.net>, "Pow
> Joshi"
> >> <pow.joshi from gmail.com> wrote:
>
> >> > so was I ....  biochem 101.... any book including Lehninger, Stryer,
> Voet
> >> >and Voet, give a detailed and lucid explanation....
> >>
> >> None of the books you mention does that ...
> >
> >well.... I remember that they did ... if we're speaking of IEF... else I
> >made a mistake.
>
> I was wrong. Just checked all three. Voet and Voet does it. Not
> in a way that's understandable by any one who does not understand
> Ohm's law but it does nevertheless. It's not what can be called
> detailed and lucid explanation but at least it's there.
>
> Stryer (the best biochemistry textbook for 101, IMHO) makes
> no mention of discontinuous electrophoresis at all.
>
> New Lehninger not only does not mention discontinuous EF but
> also does atrocious job describing EF in general and I think
> makes several errors doing it:
>
> - "Migration may also be affected by protein shape". (It's not
> "may be", it's "is").
> - "In EF, the force moving the macromolecule is the electric
> potential, E"  (wrong, driving force is an electric field).
> - "The electrophoretic mobility of the molecule, mu, is the
> ratio of the velocity of the particle molecule, V, to the electrical
> potential E. (It's "electric". Wrong. E is electric field strength).
> - "mixture of low molecular weight organic acids and bases
> (ampholytes, p.81)". (How about writing properly: mixture of
> zwitterions having different pI values that cover desired pH
> range?).
> - Why smaller popypeptides are moving more rapidly is never
> explained. The statement that precedes SDS-PAGE serves
> only to confuse: "The polyacrylamide gel acts as a molecular
> sieve, slowing migration of proteins approximately to their
> charge- to-mass ratio". (Umm, but the whole idea of SDS
> is to make approximately the same charge to mass ratio
> for all proteins!).



just a quick comment ... Correct me of I am wrong, I noticed that everyone
quotes the paper my Laemmelli et al (I think it is the 1974 paper). However,
when i went and checked out the paper, it does not discuss the technique and
the methodology in details at all; it is a paper that for the firts time
resolved some humungous number of lambda phage proteins or some such thing.
I have then found a whole bunch of literature in Methods in Enzymol etc.,
that is a post-facto  "mechanistic" viewpoint by different authors but never
a paper by Laemmelli  which discusses SDS-PAGE and the discontinuous buffer
system, as a technique in detail... would anyone know where such a paper
would exist?

thanks much
Pow


>
> DK
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