Brown D, Lydon J, McLaughlin M, Stuart-Tilley A, Tyszkowski R, Alper S
Department of Pathology, Massachusetts General Hospital, Boston, MA 02129, USA.
Histochem Cell Biol. 1996 Apr;105(4):261-7. doi: 10.1007/BF01463929.
A simple method for antigen retrieval in tissue sections and cell cultures is described. Because many antibodies recognize denatured proteins on western blots, but are poorly reactive by immunocytochemistry, the effect of applying sodium dodecyl sulfate (SDS) to cryostat sections of tissues and to cell cultures prior to immunostaining was examined. In many cases, a 5-min pretreatment with 1% SDS produced a dramatic increase in staining intensity by indirect immunofluorescence. Among the antibodies tested that showed a positive effect of SDS were an anti-Na/K-ATPase monoclonal antibody, an anti-AE1/2 anion exchanger polyclonal antipeptide antibody, a monoclonal anti-caveolin antibody, and an anti-rab4 monoclonal antibody. In other cases, including antibodies against gp330, aquaporin 1, and aquaporin 2, no effect of SDS was detected. The results show that SDS treatment can be used as a simple method of antigen retrieval in cryostat sections and on cultured cells. In some cases, antigens were not detectable without pretreatment with SDS.
本文描述了一种用于组织切片和细胞培养中抗原修复的简单方法。由于许多抗体能识别蛋白质印迹上的变性蛋白，但通过免疫细胞化学检测时反应性较差，因此研究了在免疫染色前将十二烷基硫酸钠（SDS）应用于组织冰冻切片和细胞培养物的效果。在许多情况下，用1% SDS预处理5分钟可使间接免疫荧光染色强度显著增加。在测试的显示SDS有阳性作用的抗体中，有抗Na/K - ATP酶单克隆抗体、抗AE1/2阴离子交换剂多克隆抗肽抗体、抗小窝蛋白单克隆抗体和抗rab4单克隆抗体。在其他情况下，包括针对gp330、水通道蛋白1和水通道蛋白2的抗体，未检测到SDS的作用。结果表明，SDS处理可作为冰冻切片和培养细胞中抗原修复的一种简单方法。在某些情况下，未经SDS预处理则无法检测到抗原。
