Kain S R, Mai K, Sinai P
CLONTECH Laboratories, Inc., Palo Alto, CA.
Biotechniques. 1994 Nov;17(5):982-7.
Human multiple tissue Western (MTW) blots are premade immunoblots prepared using proteins isolated from adult human tissue. The proteins are isolated from whole tissue homogenates under conditions designed to minimize proteolysis and to ensure maximal representation of tissue-specific proteins. Sodium dodecyl sulfate (SDS) solubilized proteins are fractionated by SDS polyacrylamide gel electrophoresis and electroblotted onto polyvinylidene fluoride membranes to generate blots ready for incubation with researcher-supplied antibodies. Each lane of an MTW blot contains an equivalent amount of total protein, allowing for the analysis of tissue-specific expression of a particular protein(s). The utility of MTW blots for Western blot applications was demonstrated by the detection of various cytoskeletal proteins and members of the annexin family of calcium-dependent, membrane-binding proteins. Several of these antigens were detected in separate cycles of antibody incubations using the same MTW blot. This approach is possible using a stripping procedure that allows the researcher to selectively remove both primary and secondary antibodies in a single incubation. The ease of multiple reprobings makes MTW blots both economical and convenient research tools for Western blot analyses of human tissue-specific proteins.
人类多组织蛋白质免疫印迹（MTW）是使用从成人组织中分离的蛋白质制备的预制免疫印迹。这些蛋白质是从全组织匀浆中分离出来的，分离条件旨在尽量减少蛋白水解，并确保组织特异性蛋白质的最大代表性。用十二烷基硫酸钠（SDS）溶解的蛋白质通过SDS聚丙烯酰胺凝胶电泳进行分离，然后电印迹到聚偏二氟乙烯膜上，以生成可供与研究人员提供的抗体孵育的印迹。MTW印迹的每条泳道都含有等量的总蛋白，从而能够分析特定蛋白质的组织特异性表达。通过检测各种细胞骨架蛋白和钙依赖性膜结合蛋白膜联蛋白家族的成员，证明了MTW印迹在蛋白质免疫印迹应用中的实用性。使用相同的MTW印迹，在抗体孵育的不同循环中检测到了其中几种抗原。使用一种剥离程序可以实现这种方法，该程序允许研究人员在一次孵育中选择性地去除一抗和二抗。多次重复检测的简便性使MTW印迹成为用于人类组织特异性蛋白质蛋白质免疫印迹分析的经济且方便的研究工具。
