native PAGE pH

Dr Engelbert Buxbaum via methods%40net.bio.net (by engelbert_buxbaum from hotmail.com)
Fri Mar 27 12:12:08 EST 2009


Am 25.03.2009, 10:59 Uhr, schrieb Nicola Traverso  
<ntravers from medicina.unige.it>:


> When in SDS-PAGE (discontinuous system) I use
> -tris-glycine pH 8.3 as electrophoretic buffer
> -tris-HCl pH 6.8 as stacking buffer
> -tris-HCl pH 8.8 as resolving buffer
> at which pH are proteins separated?
> I though at 8.8, i.e. the pH of the resolving buffer, but I've recently  
> read that they separate at pH 9.5! How can this be true?

The answer to this question is fairly complicated, and may be found in the  
following papers:

@article{Orn-62,
         AUTHOR= {L. Ornstein},
         TITLE= {Disk electrophoresis: {I}. {B}ackground and theory},
         YEAR= {1962},
         JOURNAL= {Ann. N. Y. Acad. Sci.},
         PAGES= {321-351},
         VOLUME= {121},
         DOI= {10.1111/j.1749-6632.1964.tb14207.x},
         LANGUAGE= {engl}
}

@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}
}


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