Specific and quantitative immunoprecipitation of tropomyosin and other cytoskeletal proteins by magnetic separation.
Specific and quantitative immunoprecipitation of tropomyosin and other cytoskeletal proteins by magnetic separation.
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通过磁力分离对原肌球蛋白和其他细胞骨架蛋白进行特异性定量免疫沉淀。
DOI：
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发表时间：
1993
期刊：
影响因子：
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通讯作者：
Fulton Ab
中科院分区：
文献类型：
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作者：
L'Ecuyer Tj;Fulton Ab
免疫沉淀是一种强大的技术，用于纯化许多蛋白质，针对这些蛋白质存在特异性抗体。磁分离最近已被证明是有效的细胞表面蛋白的免疫沉淀。我们已经使用磁性分离与抗免疫球蛋白或蛋白A结合到磁性颗粒免疫沉淀标记的肌肉原肌球蛋白和其他几种细胞骨架蛋白的特异性抗体存在。我们发现没有必要将抗原特异性抗体结合到磁性颗粒上，从而增加了该技术的通用性。使用磁性分离从肌肉培养物中定量回收原肌球蛋白上级Staph A（蛋白A阳性金黄色葡萄球菌细胞）。磁分离的特异性也优于葡萄球菌A的肌肉原肌球蛋白的免疫沉淀。成纤维细胞原肌球蛋白、波形蛋白（来自肌肉和成骨细胞）和肌球蛋白重链是其他易于用磁性分离回收的细胞骨架蛋白。因此，磁分离似乎是一种有价值的技术，从各种细胞类型的细胞骨架蛋白的免疫沉淀。
Immunoprecipitation is a powerful technique for purifying many proteins for which specific antibodies exist. Magnetic separation has recently been demonstrated to be effective in the immunoprecipitation of cell-surface proteins. We have used magnetic separation with anti-immunoglobulin or protein A bound to magnetic particles to immunoprecipitate labeled muscle tropomyosin and several other cytoskeletal proteins for which specific antibodies exist. We have not found it necessary to bind antigen-specific antibody to the magnetic particles, increasing the versatility of the technique. The quantitative recovery of tropomyosin from muscle cultures using magnetic separation is superior to Staph A (protein A-positive Staphylococcus aureus cells). The specificity of magnetic separation also compares favorably with Staph A for immunoprecipitation of muscle tropomyosin. Fibroblast tropomyosin, vimentin (from muscle and osteoblast) and myosin heavy chain are other cytoskeletal proteins that are easily recovered with magnetic separation. Magnetic separation, therefore, appears to be a valuable technique for the immunoprecipitation of cytoskeletal proteins from various cell types.