用于蛋白质复合物免疫纯化的磁珠偶联
Conjugation of magnetic beads for immunopurification of protein complexes.
作者信息
Cristea Ileana M, Chait Brian T
出版信息
Cold Spring Harb Protoc. 2011 May 1;2011(5):pdb.prot5610. doi: 10.1101/pdb.prot5610.
Abstract
Knowledge of the composition of protein complexes provides key insights into their functions. Immunoaffinity purification provides an effective means for isolating protein complexes and elucidating their composition. Immunoisolation is achieved with antibodies directed either specifically against the proteins of interest or against tags that are coupled to the proteins of interest. This approach uses immunoaffinity purification on magnetic beads coated with antibodies for the rapid and efficient purification of protein complexes from cells or tissues. This protocol describes conjugation of magnetic beads with antibodies in preparation for immunopreciptation of protein complexes.
摘要
了解蛋白质复合物的组成有助于深入了解其功能。免疫亲和纯化是分离蛋白质复合物并阐明其组成的有效方法。免疫分离是通过针对目标蛋白质或与目标蛋白质偶联的标签的特异性抗体来实现的。该方法利用包被抗体的磁珠进行免疫亲和纯化，以快速有效地从细胞或组织中纯化蛋白质复合物。本方案描述了磁珠与抗体的偶联，为蛋白质复合物的免疫沉淀做准备。
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本文引用的文献
1
Affinity purification of protein complexes.蛋白质复合物的亲和纯化。
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2
Tracking and elucidating alphavirus-host protein interactions.追踪和阐明甲病毒与宿主蛋白的相互作用。
J Biol Chem. 2006 Oct 6;281(40):30269-78. doi: 10.1074/jbc.M603980200. Epub 2006 Aug 8.
3
Fluorescent proteins as proteomic probes.作为蛋白质组学探针的荧光蛋白
Mol Cell Proteomics. 2005 Dec;4(12):1933-41. doi: 10.1074/mcp.M500227-MCP200. Epub 2005 Sep 9.
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