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- W2077801746 abstract "Reduction and alkylation of protein disulfides prior to IEF, when performed directly in a centrifugal ultrafiltration device, provides an effective means of terminating the alkylation reaction, concentrating the proteins for analysis, and removing ionic impurities that interfere with IEF. When cells were lysed in “buffers” that support the activity of enzymes such as lysozyme and benzonase, the conductivity of the resulting lysate was an order of magnitude higher than when lysis was induced by chaotropic urea detergent solutions. Following reduction and alkylation, the conductivity of both lysates was lowered by ultrafiltration to the 0.1−0.2 mS/cm range in preparation for IEF. The detergent 3-(4-heptyl)phenyl 3-hydroxypropyl dimethylammonio propanesulfonate (C7BzO), which favors the solubilization of proteins, but which interferes with SDS equilibration and second dimension PAGE, was effectively removed by ultrafiltration and exchanged with CHAPS without measurable loss of protein. Disparate protein patterns of Rhodopseudomonas palustris lysates were revealed by two-dimensional gel electrophoresis depending on which reagent was used to induce cell lysis. Keywords: acrylamide • alkylation • isoelectric focusing • pressure cycling technology • protein solubility • Rhodopseudomonas palustris • tributylphosphine • two-dimensional gel electrophoresis • ultrafiltration" @default.
- W2077801746 created "2016-06-24" @default.
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- W2077801746 date "2006-03-16" @default.
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- W2077801746 title "Simultaneous Reduction and Alkylation of Protein Disulfides in a Centrifugal Ultrafiltration Device Prior to Two-Dimensional Gel Electrophoresis" @default.
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- W2077801746 doi "https://doi.org/10.1021/pr050439w" @default.
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