关于 oct4 ssea4 tra-81-1的免疫荧光染色问题

xyzengh

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1 @+ N4 _6 ]7 Y0 p* O6 A% \; }  G/ Q% TOct4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。8 o2 S' A  k. R/ s, |3 g7 a

7 K( s: \9 J3 T, u+ r, dImmunofluorescence Analysis% _: B  k; ~$ q+ K# }$ N
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.

xyzengh

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再附上几张图让你有个感性认识吧。

xyzengh

10分钟就足够了

xyzengh

不是，普通的荧光显微镜就行了。