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A rapid assay for assessment of sphingosine kinase inhibitors and substrates
Authors:Kharel Yugesh  Mathews Thomas P  Kennedy Andrew J  Houck Joseph D  Macdonald Timothy L  Lynch Kevin R
Affiliation:aPhysical and Materials Chemistry Division, National Chemical Laboratory, Pune, Maharashtra 411008, India;bBiochemical Sciences Division, National Chemical Laboratory, Pune, Maharashtra 411008, India
Abstract:The effect of incorporating carbon nanotubes (CNTs) in the gel matrix on the electrophoretic mobility of proteins based on their molecular weight differences was investigated using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). More specifically, a reduction in standard deviation in the molecular weight calibration plots by 55% in the case of multiwalled carbon nanotubes (MWCNTs) and by 34% in the case of single-walled carbon nanotubes (SWCNTs) compared with that of pristine polyacrylamide gels was achieved after incorporating an insignificant amount of functionalized CNTs into the gel matrix. A mechanism based on a more uniform pore size distribution in CNT modified polyacrylamide gel matrix is proposed. Furthermore, the impact of SWCNTs and MWCNTs on the mobility of proteins in different molecular weight regimes at a given acrylamide concentration offers a tunable gel matrix in terms of the selection of molecular weight ranges of proteins. The robustness and excellent reproducibility of the CNT–PAGE protocol are expected to have a significant impact on the molecular weight determination of newly isolated proteins.
Keywords:Protein separation   Molecular weight determination   Gel electrophoresis   Carbon nanotubes   Polyacrylamide
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