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Electrophoretic size separation of proteins treated with sodium dodecyl sulfate in 1% agarose gels
Author(s) -
Acevedo Fernando,
Marín Viviana,
Wasserman Moisés
Publication year - 1995
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.11501601231
Subject(s) - agarose , sodium dodecyl sulfate , chromatography , chemistry , electrophoresis , gel electrophoresis , gel electrophoresis of nucleic acids , molecular weight size marker , agarose gel electrophoresis , polyacrylamide gel electrophoresis , sodium , polyacrylamide , analytical chemistry (journal) , gel electrophoresis of proteins , biochemistry , enzyme , organic chemistry , polymer chemistry , dna
Abstract Separation of proteins treated with sodium dodecyl sulfate (SDS) according to molecular size was achieved by discontinuous electrophoresis in vertical low‐concentration agarose gels. A linear relationship was found between the migration distance and the square root of the molecular weight. This holds for proteins in the range of 7–200 kDa separated in 1.25% w/v agarose gel slabs (7 × 7 × 0.15 cm) with 0.1% w/v SDS and sulfate as leading ion. The linear regression coefficient was 0.998. The molecular weight and charge of coions influenced the separation. Small ions with low p K a values were found suitable as coions. The migration distance of proteins treated with SDS varied linearly with the agarose concentration of the gel. The agarose type and quality affected the resolution of the SDS‐protein bands. We conclude that agarose gels can substitute polyacrylamide gels for the separation of proteins treated with SDS. A homogeneous agarose gel at a concentration of about 1% is a nonsieving support for electrophoresis. Therefore, the separation described here cannot be explained by the pore size of the gel. The results suggest that the separation is mainly due to the relative migration velocities of the coion and the proteins treated with SDS.