我是參考Cell Signaling Technology 03-04
裡面第396頁提到
Cell fixation有兩種方法:
1.Methnol (protein precipitation) fixation:
Immerse coverslips in 100% methnol at -20C for 10 min
2.Paraformaldehyde (cross-linking) fixation:
2a Immerse coverslips in 4% paraformaldehyde at room temp. for 10 min
2b Wash coverslips once with PBS
2c Aspirate completely and then permeabilize cells with 0.2% Triton X-100
for 5 min at room temp. or, alternatively with methnol at -20C for 5 min.
請問這兩種fixation方式究竟有何不一樣？(cross-linking and protein precipitation)
還有我的sample是sperm ，想要染出膜上或膜內的某個蛋白domain，我該fixation嗎?
最後上述2C，paraformaldehyde後，改以triton或methnol有何不同呢？
問題很多，麻煩大家，謝謝
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