Cohen B B, Moxley M, Crichton D, Deane D L, Steel C M
J Immunol Methods. 1984 Dec 14;75(1):99-105. doi: 10.1016/0022-1759(84)90229-1.
Conventional cleavage of linked polypeptide chains by heating in SDS can so alter molecular structure as to interfere with antibody binding, on which both immunoprecipitation and 'western blotting' depend. As an alternative, gentle treatment with acid at room temperature or at 0 degrees C was effective in separating the alpha and beta chains of human MHC Class II glycoprotein dimers and proved superior in terms of preservation of at least one labile epitope on the beta chain.
在十二烷基硫酸钠(SDS)中加热对连接的多肽链进行常规裂解,可能会极大地改变分子结构,从而干扰抗体结合,而免疫沉淀和“蛋白质免疫印迹法”均依赖于此。作为替代方法,在室温或0℃下用酸进行温和处理,可有效分离人主要组织相容性复合体(MHC)Ⅱ类糖蛋白二聚体的α链和β链,并且在保留β链上至少一个不稳定表位方面表现更优。