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本帖最后由 xyzengh 于 2009-9-8 22:39 编辑
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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。3 u* g1 P9 k$ @" ?- R2 o! i
' a. O3 }7 O; L$ P V) v |Immunofluorescence Analysis( X1 m4 o) O' h5 y& I' C" \" X. t2 R
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. |
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