Anti ki 67 catalog 12202 s

Staining was performed by the Vanderbilt University Medical Center Translational Pathology Shared Resource (Nashville, TN) as follows: Slides were placed on the Leica Bond Max IHC stainer. All steps besides dehydration, clearing, and coverslipping are performed on the Bond Max. Slides are deparaffinized. Heat-induced antigen retrieval was performed on the Bond Max using their Epitope Retrieval 2 solution for 20 min. Slides were placed in a Protein Block (Ref# x0909, DAKO, Carpinteria, CA) for 10 min. The sections were incubated with anti-Ki-67 (Catalog #12202 S, Cell Signaling Technology, Danvers, MA) diluted 1:250 for 1 h. The Bond Refine Polymer detection system was used for visualization. Slides were the dehydrated, cleared, and coverslipped. All images were collected on an Olympus BX41 Microscope equipped with an Olympus DP71 camera using the ×4, ×20, and ×40 objectives. Ki-67-stained area of lung lesions was quantified using the ImageJ software with manual tumor area contouring and automatic color thresholding.