Improved methodology for analysis and quantitation of proteins on one-dimensional silver-stained slab gels.

A sodium dodecyl sulfate-discontinuous polyacrylamide gel electrophoresis system for separation and quantitation of low-molecular-weight (75 to 10K Da) proteins from single muscle fibers is described. Slab gels (0.75 mm thick) were stained using an improved silver-stain technique which does not require photographic fixers in order to achieve low-level background staining. The modified staining procedure uses continuous flow washing to minimize the handling of gels. The procedure has high sensitivity and gave a linear response between approximately 2 and 70 ng of protein per band. In addition, a convenient method for mounting slab gels for photography, scanning, and long-term storage has been developed.

[1]  B. Oakley,et al.  A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. , 1980, Analytical biochemistry.

[2]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[3]  L. Adams,et al.  Ultrasensitive silver‐based color staining of polypeptides in polyacrylamide gels , 1981 .

[4]  L. Heilmeyer,et al.  Isolation, characterization and phosphorylation pattern of the troponin complexes TI2C and I2C. , 1979, European journal of biochemistry.

[5]  J. Morrissey,et al.  Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. , 1981, Analytical biochemistry.

[6]  E. McConkey,et al.  Silver Stains for proteins in polyacerylamide gels: A comparison of six methods , 1981 .

[7]  R. Moss,et al.  Physiological effects accompanying the removal of myosin LC2 from skinned skeletal muscle fibers. , 1982, The Journal of biological chemistry.

[8]  A. L. Latner,et al.  Incorporation of methylamine in an ultrasensitive silver stain for detecting protein in thick polyacrylamide gels , 1981 .

[9]  J. Potter,et al.  The content of troponin, tropomyosin, actin, and myosin in rabbit skeletal muscle myofibrils. , 1974, Archives of biochemistry and biophysics.

[10]  S. Lowey,et al.  Light Chains from Fast and Slow Muscle Myosins , 1971, Nature.

[11]  F. Julian,et al.  Mechanical properties and myosin light chain composition of skinned muscle fibres from adult and new‐born rabbits. , 1981, The Journal of physiology.

[12]  C. Merril,et al.  A rapid sensitive silver stain for polypeptides in polyacrylamide gels. , 1981, Analytical biochemistry.

[13]  C. Merril,et al.  A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels. , 1979, Analytical biochemistry.