Effect of operational variables on the separation of proteins by capillary sodium dodecyl sulfate‐gel electrophoresis

Abstract

Capillary sodium dodecyl sulfate (SDS)-gel electrophoresis, for fast and high-resolution separations of protein molecules based on their molecular masses was investigated, employing a polymeric sieving network of polyethylene oxide (PEO). A standard mixture of five proteins ranging from 14.2-66.0 kDa in molecular mass was used to evaluate the sieving matrix. A general migration velocity equation was derived for capillary SDS-gel electrophoresis of proteins and supported by the experimental data. This equation gives a better understanding of SDS-protein separations via capillary gel electrophoresis. Results are presented regarding the effects of different operational variables such as gel concentration, electric field strength, molecular mass, and temperature on the electrophoretic migration properties of the different size protein molecules.

Ultrathin-layer sodium dodecyl sulfate gel electrophoresis of proteins: Effects of gel composition and temperature on the separation of sodium dodecyl sulfate-protein complexes

“…The number of SDS molecules in the SDS-protein complex is proportional to the M r and the exponent k represents information about the apparent shape of the SDS-covered polypeptide chain during its migration due to the influence of the applied electric field [24]. The separation of SDS protein complexes is an activated process, which requires a certain amount of activation energy (E a ) for passage through the viscous separation media.…”

Activation energy associated with the electromigration of oligosaccharides through viscosity modifier and polymeric additive containing background electrolytes

“…CE has been extensively used for physicochemical characterization of biopolymers, such as nucleic acids, proteins, and carbohydrates using cross‐linked gels or linear polymer matrices . Based on the fact that the separation mechanism of biopolymers in linear polymer networks are similar to that of in cross‐linked gels, and due to the difficulties of working with chemical gels in narrow bore capillary tubings, today almost exclusively physical gels are used in CE.…”

Subnanomolar detection limit for sodium dodecyl sulfate — capillary gel electrophoresis using a fluorogenic, noncovalent dye

“…For improved resolution, a higher percentage of polyacrylamide or a step gradient [ll] is suggested. Capillary thermostating at some optimal temperature has also been shown to increase peak efficiency significantly [18]. Separation of standard proteins in the molecular mass range of 29 kDa-78 kDa was complete within 18 min.…”