Abstract:
Immunoblotting, also termed western blotting, is a powerful method for detection and characterization of proteins separated by various electrophoretic techniques. The combination of sodium dodecyl sulfate-poly acrylamide gel electrophoresis (SDS-PAGE), having high separating power, immunoblotting to synthetic membranes, and detection with highly specific peptide antibodies, is especially useful for studying individual proteins in relation to cellular processes, disease mechanisms, etc. Here, we describe a protocol for the sequential detection of various forms of an individual protein using peptide antibodies, exemplified by the characterization of antibody specificity for different forms of the protein calreticulin by double SDS-PAGE immunoblotting.
摘要:
免疫印迹,也被称为西方印迹,是通过各种电泳技术分离的蛋白质的检测和表征的强大方法。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),具有高分离能力,免疫印迹合成膜,用高度特异性的肽抗体检测,对于研究与细胞过程相关的单个蛋白质特别有用,疾病机制,等。这里,我们描述了使用肽抗体顺序检测各种形式的单个蛋白质的方案,通过双重SDS-PAGE免疫印迹表征不同形式的蛋白钙网蛋白的抗体特异性。