Sodium dodecyl sulfate-capillary gel electrophoresis of proteins using non-cross-linked polyacrylamide.
Proteins with relative molecular masses of 14,000 to 205,000 were separated by sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) using non-cross-linked linear polyacrylamide gels on both coated and uncoated fused-silica capillaries. It was determined that viscosity of the acrylamide solution was a major factor affecting column stability with linear acrylamide gels. When the viscosity of the acrylamide solution reaches 100 cP, electro-osmotically driven displacement of the gels is insignificant. Uncoated capillaries provided better resolution, stability, and reproducibility than surface coated capillaries when the concentration of linear polyacrylamide was greater than 4%. At lower gel concentrations, non-cross-linked polyacrylamide is easily displaced from the columns. A calibration plot of log molecular mass vs. mobility with non-linear polyacrylamide was linear, which indicated that resolution was equivalent to that obtained with cross-linked acrylamide. Separations with model proteins indicated that baseline resolution between protein species that vary 10% in molecular mass can be achieved.
Wu
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Department of Chemistry, Purdue University, West Lafayette, IN 47907.
Regnier
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D000180:Acrylic Resins / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D008970:Molecular Weight; D009211:Myoglobin / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D014783:Viscosity; D000099166:Polyacrylamides
D000180:Acrylic Resins; D007074:Immunoglobulin G; D009211:Myoglobin; D011506:Proteins; D000099166:Polyacrylamides