Ki67 and CD-31 IHC of tumor sections was performed as previously described [14 (link)]. Paraffin sections (4-μm-thick) of the biggest tumor sections from ID8, ID8-c-MYC, and ID8-KRAS mice (sections were taken when BW exceeded 23 g) were dewaxed in xylene and rehydrated through graded ethanol to water. Antigens were retrieved by boiling in 10 mM citrate buffer (pH 6.0) for 30 min. The cooled sections were incubated in DAKO REAL Peroxidase-Blocking solution (DAKO, Carpinteria, CA, USA) for 10 min to quench endogenous peroxidase. Sections were incubated in DAKO Protein Blocking solution (DAKO) at room temperature for 10 min to block non-specific binding. Sections were then stained for Ki67 using rabbit monoclonal antibody against mouse Ki67(1:100; Spring Bioscience, CA, USA), CD-31 using a rat monoclonal antibody against mouse CD-31 (ab56299, Abcam, Tokyo, Japan, 1:100 dilution).
Free full text: Click here
