can i use EDTA instead of TEMED for SDS PAGE ?
Sudheendra Rao N R
(by sudhee26 from gmail.com)
Fri May 30 19:53:32 EST 2008
I see the discusssion to basics..its very interesting
I got this while surfing..any use is welcome
also i have seen EMBL webpage about 3D gel electrophoresis...hahaha..1D-SDS
PAGE itself has somany people still searching for its biophysics...3D is far
On Sat, May 31, 2008 at 2:12 AM, Pow Joshi <pow.joshi from gmail.com> wrote:
> 2008/5/29 DK <dk from no.email.thankstospam.net>:
> > In article <mailman.251.1212073353.3533.methods from net.bio.net>, "Pow
> > <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).
> when i went and checked out the paper, it does not discuss the technique
> 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
> 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
> > DK
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