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Author:
Date:
2013-08-05
[Abstract] This protocol was developed to label proteins in bacterial cells with antibodies conjugated to a fluorophore for fluorescence microscopy imaging. The procedure is optimized to minimize morphological changes and also to minimize the amount of antibodies needed for the staining. The protocol can also be used with primary antibodies conjugated to a fluorophore. The method has been verified extensively (van der Ploeg et al., 2013), but it should be noted that one case in Caulobacter crescentus (Hocking et al., 2012) has been reported in which the localization of a protein changed upon fixation by formaldehyde/glutaraldehyde. However, the localization of the same protein in E. coli did not change.
[摘要] 该协议被开发来标记细菌细胞中的蛋白质与缀合到荧光显微镜成像荧光的抗体。 优化程序以使形态变化最小化，并且使染色所需的抗体的量最小化。 该方案还可以与结合至荧光团的一抗一起使用。 该方法已经广泛地验证（van der Ploeg等人，2013），但是应当注意的是，在新月柄杆菌中的一种情况（Hocking等人，/2012>），其中蛋白质的定位在通过甲醛/戊二醛固定时改变。 然而，相同蛋白质在E中的定位。 大肠杆菌没有改变。
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