Recombinant Anti-CD34 antibody [EP373Y] (ab81289)

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Publication protocol

The liver tissues embedded in paraffin were sliced into 4–6 mm pieces, deparaffinized and serially dehydrated in ethanol. The sections were boiled for 3 minutes while immersed in EDTA-Tris (pH 9.0) for antigen retrieval and then treated with hydrogen peroxide for 30 minutes to inactivate the endogenous peroxidase. The sections were incubated overnight at 4 °C with primary antibodies, which included antibodies against α-SMA (1:400; Abcam, Cambridge, MA, USA), CD31 (1:2000; Abcam), CD34 (1:2500; Abcam) and von Willebrand factor (VWF) (1:200; Proteintech, Wuhan, China). The sections were subsequently incubated with the appropriate biotinylated secondary (goat anti-rabbit IgG and goat anti-mouse IgG; Origene Technologies, Beijing, China) for 30 minutes at room temperature, stained with Diaminobenzidine (DAB) and counterstained with hematoxylin. The positive areas were colored a brownish yellow. Image-Pro Plus software 6.0 was used for the immunohistochemical analysis.

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