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Partitioning of Proteins in Triton X-114

  • James G. Pryde
Part of the Methods in Molecular Biology™ book series (MIMB, volume 88)

Abstract

Cultured mammalian cells and tissues can be solubilized by the nonionic detergent Triton X-114 (octylphenoxy polyethoxyethanol). This detergent, when in solution above its critical micelle concentration, increases its micelle weight when warmed from 0 to 20°C and in the process decreases its critical micelle concentration. This induces intermicellar interactions, which lead to turbidity (the cloud point) and phase separation of the detergent at 20°C. A simple low-speed centrifugation step recovers the detergent-enriched phase as an oily pellet. Following solubilization and warming to 20°C, integral membrane proteins partition into the detergent-enriched phase, and peripheral and cytosolic proteins are recovered from the detergent-depleted aqueous phase.

Keywords

Critical Micelle Concentration Integral Membrane Protein Chromaffin Granule Immunoprecipitation Buffer Cobalt Nitrate Hexahydrate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc. 1998

Authors and Affiliations

  • James G. Pryde
    • 1
  1. 1.Respiratory Medicine Unit, Rayne Laboratory, Department of Medicine (RIE)University of Edinburgh Medical SchoolEdinburghUK

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