can i use EDTA instead of TEMED for SDS PAGE ?

Dr. Hiranya S. Roychowdhury via methods%40net.bio.net (by hroychow from nmsu.edu)
Wed Jun 4 09:13:14 EST 2008


Thanks for rsponding to this email, Engelbert.  I am out of town at the
moment and had not had a chance to check my emails regularly.

Hiranya


> Am 30.05.2008, 16:35 Uhr, schrieb WS <novalidaddress from nurfuerspam.de>:
>
>> Dear Hiranya,
>>
>> is there any chance to obtain this paper? I am (sort of desperately)
>> looking for literature which really explains what is the chemistry and
>> physics behind the different gel systems (Tris-Glycine, Tris-Tricine,
>> how stacking gels work, what improves and what worsens the resolution
>> etc.).
>>
>> Are there any other systems that add copolymers to acrylamide or use
>> something completely different? (You can see, I really want to mess it
>> up...)
>
> For the theory, look at the following papers:
>
> @article{Jov-73a,
>          AUTHOR= {T.M. Jovin},
>          TITLE= {Multiphasic Zone Electrophoresis. {I}. {S}teady-State
> Moving-Boundary Systems Formed by Different Electrolyte Combinations},
>          JOURNAL= {Biochemistry},
>          VOLUME= {12},
>          YEAR= {1973},
>          PAGES= {871-879},
>          LANGUAGE= {engl}
> }
>
> @article{Jov-73b,
>          AUTHOR= {T.M. Jovin},
>          TITLE= {Multiphasic Zone Electrophoresis. {II}. Design of
> Integrated Discontinuous Buffer Systems for Analytical and Preparative
> Fractionation},
>          JOURNAL= {Biochemistry},
>          VOLUME= {12},
>          YEAR= {1973},
>          PAGES= {879-890},
>          LANGUAGE= {engl}
> }
>
> @article{Jov-73c,
>          AUTHOR= {T.M. Jovin},
>          TITLE= {Multiphasic Zone Electrophoresis. {III}. {F}urther
> Analysis and New Forms of Discontinuous Buffer Systems},
>          JOURNAL= {Biochemistry},
>          VOLUME= {12},
>          YEAR= {1973},
>          PAGES= {890-898},
>          LANGUAGE= {engl}
> }
>
> @article{Jov-73d,
>          AUTHOR= {T.M. Jovin},
>          TITLE= {Multiphasic zone electrophoresis. {IV} {D}esign and
> analysis of discontinuous buffer systems with a digital computer},
>          JOURNAL= {Ann. N.Y. Acad. Sci.},
>          VOLUME= {209},
>          YEAR= {1973},
>          PAGES= {477-496},
>          LANGUAGE= {engl}
> }
>
> There is a web site for the calculation of buffer systems satisfying
> certain criteria at
> http://www.buffers.nichd.nih.gov/, but it currently seems non-operational.
>
> You can of cdourse use different detergents, in particular those with
> positive charge. As I explained in the following paper they have
> advantages with certain types of proteins:
>
> @article{Bux-03,
>          AUTHOR= {E. Buxbaum},
>          TITLE= {Cationic Electrophoresis and Electrotransfer of Membrane
> Glycoproteins},
>          YEAR= {2003},
>          JOURNAL= {Anal. Biochem.},
>          PAGES= {70-76},
>          VOLUME= {314},
>          LANGUAGE= {engl},
>          DOI= {10.1016/S0003-2697(02)00639-5}
> }
>
> In theory one could also link affinity tags to the matrix to get an
> affinity electrophoresis in analogy of affinity chromatography, but I am
> not aware of anybody ever having done that. Other than that the matrix
> only has to provide the right pore size, its chemical composition is not
> relevant. So starch or agar gels are used for some problems, but only
> because they offer larger pores than acrylamide.
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>


-- 
Hiranya S. Roychowdhury, Ph.D.
Asst. Professor,
Health & Public Services
Dona Ana Community College
New Mexico State University
Las Cruces, NM 88003



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