本帖最后由 xyzengh 于 2009-9-8 22:39 编辑 ) y0 N/ ~, P1 U' C* Z
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Oct4是核抗原，所以triton是必须的，SSEA3/4是膜表面抗原，可加可不加，你为了方便可以都加，把我文章里面的方法发上来给你参考吧。. N8 W6 W8 H- H, G( E. z1 a! J
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Immunofluorescence Analysis; r/ A4 M; B; T6 Q% a6 k
Cells were fixed with 4% paraformaldehyde for 10 min., incubated in PBS containing 5% rabbit serum and 0.4% Triton X-100 for blocking and permeablization, respectively. PBS containing 0.5% Tween 20 (PBS-T) was used to dilute antibodies and wash the cells in the following procedures. The cells were incubated with antibodies against OCT4, SSEA-4，TRA-1-60 and TRA-1-81 at 37 °C for 1 h, followed by washing with PBS-T for three times. Afterwards, the cells were incubated with fluorochrome-conjugated, corresponding secondary antibodies at 37 °C for 45 min. and washed with PBS-T for three times. Finally, the cells were examined under fluorescence microscope to capture both phase and fluorescent images.作者: xyzengh 时间: 2009-9-8 22:38