shanghainese

CD31应该在细胞膜上。
你的 isotype control 呢？

shanghainese

TLD-3A12，你用的是这个克隆吗？
The TLD-3A12 antibody is tested for immunohistochemical staining of acetone-fixed frozen sections and zinc-fixed
paraffin sections.
它是不能用在多聚甲醛固定的组织细胞上的。

shanghainese

用丙酮固定.

shanghainese

引用:

原帖由 悠悠我心 于 2012-5-9 18:56 发表
我做过冰冻片，多聚甲醛固定后再修复可以做出来。不修复就做不出来。

冰冻片丙酮固定,就不需要修复这一步骤了.

shanghainese

引用:

原帖由 摞贝 于 2012-5-14 15:23 发表
您好，我做的是细胞爬片   可以用丙酮固定吗？  您做的是石蜡切片还是细胞爬片呢？

细胞爬片当然可以丙酮固定。

shanghainese

I) Culture cells to 50-70% confluency. Some cells (endothelial/epithelial) may require a higher level of confluency. For suspension cells in a regular petri dish or flask, transfer enough cells suspended in medium to each well of a poly-D-lysine-coated microslide. Then Incubate microslide in CO2 incubator from 1 hour to overnight to allow cells to attach to the bottom of the slide. The incubation time varies depending on the suspension cell used. Check for cell attachment using a microscope.

Gently aspirate medium from the wells, wash the attached cells briefly with 1x PBS and dry cells.

II) Fix slides in cold acetone (-20oC) for 10 minutes, dry again and followed by 2 PBS washes.

shanghainese

不用客气。