从培养的哺乳动物细胞中制备耐去污剂膜（DRMs）。

Preparation of detergent-resistant membranes (DRMs) from cultured mammalian cells.

作者信息

Brown Deborah A

机构信息

Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, NY, 11794-5215, USA,

出版信息

Methods Mol Biol. 2015;1232:55-64. doi: 10.1007/978-1-4939-1752-5_5.

DOI:10.1007/978-1-4939-1752-5_5

PMID:25331127

Abstract

Detergent-resistant membranes (DRMs) isolated from cells are enriched in proteins and lipids with a high affinity for lipid rafts, or membrane microdomains in the liquid-ordered phase. Enrichment in DRMs provides a good indication that a protein is "raftophilic," and may be present in rafts in cell membranes before extraction. Here, I describe preparation of Triton X-100-insoluble DRMs from cultured mammalian cells on sucrose gradients. Methods for analyzing the distribution of particular proteins across the gradient, and for recovering DRMs for further use are presented.

摘要

从细胞中分离出的抗去污剂膜（DRM）富含对脂筏（即处于液相有序相的膜微区）具有高亲和力的蛋白质和脂质。DRM中的富集充分表明一种蛋白质是“嗜筏的”，并且在提取之前可能存在于细胞膜的脂筏中。在这里，我描述了在蔗糖梯度上从培养的哺乳动物细胞中制备Triton X-100不溶性DRM的方法。还介绍了分析特定蛋白质在梯度中的分布以及回收DRM以供进一步使用的方法。