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.
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© 1998 Humana Press Inc.
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Pryde, J.G. (1998). Partitioning of Proteins in Triton X-114. In: Clegg, R.A. (eds) Protein Targeting Protocols. Methods in Molecular Biology™, vol 88. Humana Press. https://doi.org/10.1385/0-89603-487-9:23
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DOI: https://doi.org/10.1385/0-89603-487-9:23
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