G8.8 anti-EpCAM

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Publication protocol

Mouse and human liver samples were fixed and paraffin-embedded for immunostaining. Then, 4 μm thick tissue sections were rehydrated and then stained with hematoxylin and eosin (HE) for histological analysis. For immuno-histochemistry, antigen retrieval was performed in citrate buffer [10 mmol/L, 0.01% Tween20, pH 6.0 for EpCAM or Tris-EDTA buffer (10 mmol/L Tris base, 1 mmol/L EDTA solution, pH 9.0 for CK19)], and for the removal of endogenous peroxidase, the samples were treated with 3% hydrogen peroxide. The sections were then incubated overnight at 4 °C in blocking solution containing primary antibodies. Blocking solution without primary antibodies only was used as a control. After undergoing appropriate secondary antibody incubation, the immunostaining was completed using a Vectastain ABC Kit (Vector Laboratories, Burlingame, CA, United States) and liquid 3,3′-diaminobenzidine (Dako). The results were analyzed using a Nikon microscope, and images were captured with NIS-Elements F4.0.

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