Anti cleaved caspase 3 asp175

Briefly, tumor tissue samples from all the different treatment groups were fixed in 10% neutral-buffered formalin for 24 h and embedded in paraffin for histology and IHC. Sections (5-μM-thick) were incubated with anti-PECAM-1 antibody (M-20) (1:200, #sc1506, Santa Cruz Biotechnology) and anti-cleaved Caspase-3 (Asp175) (1:300, #sc7272, Santa Cruz Biotechnology) at 4 °C overnight. Negative controls were carried out by omitting the primary antibodies. Immunostaining was accomplished using a Benchmark immunostainer (Ventana, Tuscon, AZ). Immunoreaction was displayed using the avidin–biotin–peroxidase complex (ABC) method. Peroxidase activity was visualized with diaminobenzidine. Counterstaining was performed with haematoxylin. Quantification of caspase 3 positive cells and CD31 was performed as previously described [21 (link),27 (link)].