Kurien Biji T, Scofield R Hal
Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, 73104, USA,
Methods Mol Biol. 2015;1312:77-86. doi: 10.1007/978-1-4939-2694-7_11.
Western blotting enables the detection and characterization of proteins of low abundance. Sodium dodecyl sulfate (SDS) polyacrylamide gel-separated proteins are normally transferred electrophoretically to nitrocellulose or polyvinylidene difluoride membranes. Here we describe the transfer proteins [Ro 60 (or SSA) autoantigen, 220 and 240 kDa spectrin antigens, and prestained molecular weight standards] by diffusion from SDS polyacrylamide gels at 37 °C. Up to 12 immunoblots can be obtained from a single gel by this method.
蛋白质免疫印迹法能够检测和鉴定低丰度蛋白质。十二烷基硫酸钠（SDS）聚丙烯酰胺凝胶分离的蛋白质通常通过电泳转移至硝酸纤维素或聚偏二氟乙烯膜上。在此，我们描述了在37℃下通过从SDS聚丙烯酰胺凝胶扩散转移蛋白质[Ro 60（或SSA）自身抗原、220和240 kDa血影蛋白抗原以及预染分子量标准品]的方法。通过这种方法，从一块凝胶中最多可获得12张免疫印迹。
