PCNA Antibody (PC10): sc-56

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Publication protocol

Mice ectopic lesions were fixed with 10% neutral buffered formalin for 24 h at room temperature, followed by immersion in 70% ethanol overnight at 4°C, then embedded in paraffin and sectioned at 4-μm thickness. For each paraffin-embedded tissue block, slide was stained by hematoxylin and eosin (H&E) and immunostained for different antibodies. For antigen retrieval, the slides were heated at 98°C in the EDTA buffer (pH 8.0) or the citric acid solution (pH 6.0) for a total of 30 min and cooled naturally at room temperature. The slides were incubated with primary antibodies against PCNA, VEGF, and E-cadherin, diluted to 1:2,000, 1:50, and 1:100 respectively at 4°C for 12 h. Further, after sufficient PBS wash, the slides were incubated with HRP-conjugated goat polyclonal antibody. The number and intensity of positive cells were counted by Image Pro-Plus 6.0 (Media Cybernetics, Inc., MD, U.S.A.). Images were obtained with the microscope fitted with a digital camera (Olympus, SD, U.S.A.).

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