Bulinski J C, Kumar S, Titani K, Hauschka S D
Proc Natl Acad Sci U S A. 1983 Mar;80(6):1506-10. doi: 10.1073/pnas.80.6.1506.
The NH2-terminal peptide of skeletal muscle alpha-actin (S alpha N peptide), which contains a primary sequence unique to this actin isozyme, was used to prepare an isozyme-specific peptide antibody. S alpha N peptide was purified from chicken breast muscle actin by preparative reverse-phase HPLC and was coupled to hemocyanin. This complex was used to immunize rabbits in order to elicit actin antibodies specific for the skeletal muscle alpha-actin isozyme. The antibody obtained, called S alpha N antibody, was reactive with S alpha N peptide and with skeletal muscle alpha-actin as well as with cardiac muscle alpha-actin. S alpha N antibody did not react with either of the actin isozymes present in smooth muscle (smooth muscle alpha and gamma) or in brain (nonmuscle beta and gamma). S alpha N antibody was used to detect muscle-specific actin in differentiating mouse and human myoblasts by using immunoblots of myoblast extracts and immunofluorescent staining of fixed cells.
骨骼肌α-肌动蛋白的氨基末端肽(SαN肽)含有该肌动蛋白同工酶特有的一级序列,用于制备同工酶特异性肽抗体。通过制备型反相高效液相色谱从鸡胸肌肌动蛋白中纯化出SαN肽,并将其与血蓝蛋白偶联。该复合物用于免疫兔子,以引发对骨骼肌α-肌动蛋白同工酶特异的肌动蛋白抗体。获得的抗体称为SαN抗体,它与SαN肽、骨骼肌α-肌动蛋白以及心肌α-肌动蛋白都有反应。SαN抗体与平滑肌(平滑肌α和γ)或脑中存在的任何一种肌动蛋白同工酶(非肌肉β和γ)都不发生反应。通过使用成肌细胞提取物的免疫印迹和固定细胞的免疫荧光染色,SαN抗体被用于检测分化中的小鼠和人类成肌细胞中的肌肉特异性肌动蛋白。