Hnasko Thomas S, Hnasko Robert M
Department of Neurosciences, School of Medicine, University of California San Diego (UCSD), 9500 Gilman Drive (BRF2), La Jolla, CA, 92093, USA,
Methods Mol Biol. 2015;1318:87-96. doi: 10.1007/978-1-4939-2742-5_9.
Western blotting is a technique that involves the separation of proteins by gel electrophoresis, their blotting or transfer to a membrane, and selective immunodetection of an immobilized antigen. This is an important and routine method for protein analysis that depends on the specificity of antibody-antigen interaction and is useful for the qualitative or semiquantitative identification of specific proteins and their molecular weight from a complex mixture. This chapter will outline the requisite steps including gel electrophoresis of a protein sample, transfer of protein from a gel to a membrane support, and immunodetection of a target antigen.
蛋白质印迹法是一种通过凝胶电泳分离蛋白质、将其印迹或转移至膜上以及对固定化抗原进行选择性免疫检测的技术。这是蛋白质分析的一种重要常规方法,它依赖于抗体 - 抗原相互作用的特异性,可用于从复杂混合物中定性或半定量鉴定特定蛋白质及其分子量。本章将概述必要的步骤,包括蛋白质样品的凝胶电泳、蛋白质从凝胶转移至膜支持物以及目标抗原的免疫检测。