Anti ki 67 rm 9106

Dissected intestinal tissues were fixed with 4% paraformaldehyde, embedded in paraffin wax and sectioned at a thickness of 4 µm. These sections were stained with H&E or were processed for further immunostaining as described previously (Oshima et al., 1995b (link)). Rabbit polyclonal anti-cleaved caspase3 (Asp175, #9661; Cell Signaling Technology), anti-Ki67 (RM-9106; Thermo Fisher Scientifics) were used as primary antibodies.

The whole mouse brains of the control and treatment groups were post-fixed in 4% paraformaldehyde for 24 h, followed by continuous dehydration in 20% and 30% sucrose solutions (24 h each) until the whole brains sank to the bottom of the solution. The frozen GBM tissues were serially sectioned into brain slices with a 12 µm thickness using a cryostat. The brain slices were photographed under a microscope. The brain tumor slices were then incubated with PBS containing 0.3% Triton X-100 at room temperature for 30 min, followed by direct blocking in 10% goat serum for 1 h. The brain tumor slices were subsequently incubated with anti-Ki67 (RM-9106, Thermo Scientific, 1:400) overnight and with secondary antibodies for an hour on the next day. DAPI dye was used for nuclear staining. The immunofluorescent signals were observed and photographed under an inverted fluorescence microscope.