can i use EDTA instead of TEMED for SDS PAGE ?

Dr Engelbert Buxbaum via methods%40net.bio.net (by engelbert_buxbaum from hotmail.com)
Tue Jun 3 10:01:37 EST 2008


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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