Optimizing Conditions for Capillary Isoelectric Focusing Separations Using
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Native capillary isoelectric focusing for the separation of protein complex isoforms and subcomplexes.
Here we report the use of capillary isoelectric focusing under native conditions for the separation of protein complex isoforms and subcomplexes. Using biologically relevant HIS-tag and FLAG-tag purified protein complexes, we demonstrate the separations of protein complex isoforms of the mammalian target of rapamycin complex (mTORC1 and 2) and the subcomplexes and different phosphorylation stat...
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We employed a liquid core waveguide to image both DNA electrophoresis separations and isoelectric focusing of proteins. The utility of the system is demonstrated for DNA fragment sizing and protein separations. The system utilizes the liquid-core waveguide as an efficient window for both the excitation of separated samples and the collection of light through total internal reflectance, with an ...
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Capillary electrophoresis (CE) is a high-resolution technique for the separation of a wide variety of molecules of biological interest such as metabolites, drugs, amino acids, nucleic acids, and carbohydrates. This unit focuses on the use of CE to separate proteins and peptides based on charge-to-mass ratios. Separation of proteins based on their isoelectric points is described along with a pro...
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Despite the significant advantages of capillary isoelectric focusing over gel isoelectric focusing, widespread acceptance of the technique as a replacement for gel isoelectric focusing has not occurred. Several factors contribute to capillary isoelectric focusing’s slower-than-expected adoption; perhaps the most important are the performance and procedural difficulties introduced when capillary...
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Capillary electrophoresis (CE) techniques have been fundamental in advancing technology for the separation and identification of peptide and protein mixtures [1,2]. Today, they have become a staple in conjunction with mass spectrometry systems to resolve, identify and analyze protein and peptide characteristics with sensitivity and accuracy. Recent advances have miniaturized traditional CE inst...
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