Brown Deborah A
Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, NY, 11794-5215, USA,
Methods Mol Biol. 2015;1232:55-64. doi: 10.1007/978-1-4939-1752-5_5.
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.
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。
