用于分离人骨骼肌肌球蛋白重链亚型的增强型蛋白质电泳技术。
Enhanced protein electrophoresis technique for separating human skeletal muscle myosin heavy chain isoforms.
作者信息
Bamman M M, Clarke M S, Talmadge R J, Feeback D L
机构信息
Department of Human Studies, University of Alabama at Birmingham 35294-1250, USA.
出版信息
Electrophoresis. 1999 Mar;20(3):466-8. doi: 10.1002/(SICI)1522-2683(19990301)20:3<466::AID-ELPS466>3.0.CO;2-7.
Talmadge and Roy (J. Appl. Physiol. 1993, 75, 2337-2340) previously established a sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) protocol for separating all four rat skeletal muscle myosin heavy chain (MHC) isoforms (MHC I, IIa, IIx, IIb); however, when applied to human muscle, the type II MHC isoforms (Ila, IIx) are not clearly distinguished. In this brief paper we describe a modification of the SDS-PAGE protocol which yields distinct and consistent separation of all three adult human MHC isoforms (MHC I, IIa, IIx) in a minigel system. MHC specificity of each band was confirmed by Western blot using three monoclonal IgG antibodies (mAbs) immunoreactive against MHCI (mAb MHCs, Novacastra Laboratories), MHCI+IIa (mAb BF-35), and MHCIIa+IIx (mAb SC-71). Results provide a valuable SDS-PAGE minigel technique for separating MHC isoforms in human muscle without the difficult task of casting gradient gels.
塔尔梅奇和罗伊（《应用生理学杂志》，1993年，75卷，2337 - 2340页）之前建立了一种十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）方法，用于分离大鼠骨骼肌的所有四种肌球蛋白重链（MHC）同工型（MHC I、IIa、IIx、IIb）；然而，当应用于人类肌肉时，II型MHC同工型（IIa、IIx）无法清晰区分。在这篇简短的论文中，我们描述了对SDS - PAGE方法的一种改进，该改进能在微型凝胶系统中对所有三种成人MHC同工型（MHC I、IIa、IIx）进行清晰且一致的分离。通过使用三种针对MHC I（mAb MHCs，诺瓦卡斯托拉实验室）、MHC I + IIa（mAb BF - 35）和MHC IIa + IIx（mAb SC - 71）具有免疫反应性的单克隆IgG抗体（mAb）进行蛋白质印迹法，证实了每条带的MHC特异性。结果提供了一种有价值的SDS - PAGE微型凝胶技术，用于分离人类肌肉中的MHC同工型，而无需进行制备梯度凝胶这项困难的工作。
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