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High‐resolution separation of peptides by sodium dodecyl sulfate‐polyacrylamide gel “focusing”
Author(s) -
Zilberstein Gleb,
Korol Leonid,
Shlar Ilya,
Righetti Pier Giorgio,
Bukshpan Shmuel
Publication year - 2008
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200700625
Subject(s) - sodium dodecyl sulfate , peptide , polyacrylamide , chemistry , polyacrylamide gel electrophoresis , chromatography , resolution (logic) , micelle , analytical chemistry (journal) , crystallography , polymer chemistry , organic chemistry , aqueous solution , biochemistry , computer science , artificial intelligence , enzyme
As a follow‐up of our previous report ( Anal. Chem. 2007, 79 , 821–827) on analytical SDS‐PAGE focusing, a refinement of the method for separation of peptides in the small to medium M r range (0.5–10 kDa) is here reported, based on a shallow gradient of immobilized positive charges (0–10 mM) onto a minimally sieving polyacrylamide gel matrix (4%T, 2.5%C). Unlike conventional SDS‐PAGE, which rarely can achieve the separations of polypeptide chains below a critical value of 10 kDa, the present method can be fine‐tuned to perform such separations even down to a size of only 500 Da. In the case of larger fragments, the major peptide zones are shown, under microscope observation, to be composed by envelops of bands as narrow as 20–100 μm, spaced at regular intervals of 100–150 μm. It is hypothesized that such larger peptides could form complexes with rather small SDS micelles and that such peptide–SDS complexes could differ in charge by just a single negative charge.