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Gradient polyacrylamide gel electrophoresis in presence of sodium dodecyl sulfate: A practical approach to muscle contractile and regulatory proteins
Author(s) -
Sobieszek Apolinary
Publication year - 1994
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.11501501151
Subject(s) - acrylamide , polyacrylamide , sodium dodecyl sulfate , glycerol , polyacrylamide gel electrophoresis , gel electrophoresis , chromatography , chemistry , sodium , electrophoresis , biochemistry , polymer , enzyme , polymer chemistry , organic chemistry , copolymer
Abstract Two gradient systems for polyacrylamide gel electrophoresis (PAGE) in the presence of sodium dodecyl sulfate (SDS) are described, with emphasis on improvements accumulated over two decades of studies on contractile proteins and regulatory enzymes from smooth muscle. The first “big slab” system utilizes 18 × 20 × 0.1 cm 3 gels and a 10–18% acrylamide gradient, optimized for a high resolution of 10 to 500 kDa polypeptides. Eight (or more) gels are cast simultaneously with a gradient formation from “bottom to top” and 20% glycerol is added to the 18% acrylamide solution. The second “minislab” system represents an improved version of the system of Matsudaira and Burgess ( Anal. Biochem. 1978, 87 , 386–396), with 8 × 10 × 0.05 cm 3 gels and 5–15% or 9–18% acrylamide gradient ranges. They are cast from “top to bottom” in 28‐piece batches also with the addition of glycerol for improved gradient formation. Both types of gels can also be cast individually using a specially designed pestle‐type gradient maker. For gel destaining, a convenient continuous hydrodynamic destainer is also described.