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Author:
Date:
2013-08-20
[Abstract] A great way to specifically isolate and quantify proteins in the cell surface membrane is to take advantage of the biotinylation technique. It consists of labeling cell surface proteins with a biotin reagent before lysing the cells, and isolating these tagged proteins by NeutrAvidin pull-down. Then, the samples are subjected to SDS-PAGE separation, transferred to PVDF membranes and probed with specific antibodies. Quantification of cell surface expression is accomplished by densitometric measurement of the bands corresponding to the protein of interest and subsequent normalization by a membrane protein (as control).
[摘要] 特异性分离和定量细胞表面膜中的蛋白质的好方法是利用生物素化技术。 它包括在裂解细胞之前用生物素试剂标记细胞表面蛋白，并通过NeutrAvidin下拉分离这些标记的蛋白。 然后，将样品进行SDS-PAGE分离，转移至PVDF膜并用特异性抗体探测。 细胞表面表达的定量通过密度测量对应于感兴趣的蛋白质的条带并随后通过膜蛋白质（作为对照）标准化来完成。
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