Immunostaining of Liver Tissue Sections

Liver tissues were fixed overnight in 4% paraformaldehyde, and then paraffin-sectioned (Penn Digestive Diseases Center Morphology Core). Immunohistochemistry was performed using standard protocols as described previously(27 (link), 28 (link)). For FAH staining we employed rabbit anti-FAH (ab81087, Abcam), and goat anti-rabbit for secondary antibody (Vector Labs). The dCas9 has a hemagglutinin (HA) tag, therefore we could stain tissues with a mouse anti-HA antibody (H3663, Sigma-Aldrich), and rabbit anti-mouse IgG1 secondary antibody (SAB3701173, Sigma-Aldrich). For MYC we used a rabbit anti-MYC antibody (sc764, Santa Cruz Biotechnology), and goat anti-rabbit as the secondary antibody (Vector Labs). Staining was performed similarly using antibodies to YAP1 (4912, Cell Signaling Technologies), AFP (sc8108, Santa Cruz Biotechnology), OPN (AF808, R&D Systems), and Ki67 (RM9106, Thermo Scientific).

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Wangensteen K.J., Wang Y.J., Dou Z., Wang A.W., Mosleh-Shirazi E., Horlbeck M.A., Gilbert L.A., Weissman J.S., Berger S.L, & Kaestner K.H. (2018). Combinatorial genetics in liver repopulation and carcinogenesis with a novel in vivo CRISPR activation platform. Hepatology (Baltimore, Md.), 68(2), 663-676.

Publication 2018

ab81087 H3663 sc764 sc8108 AF808 RM9106

Anti antibody Antibodies Digestive diseases Goat Hemagglutinin Igg1 Immunohistochemistry Liver Mouse Paraffin Paraformaldehyde Rabbit Stain Tissues Vector Yap1

Corresponding Organization : University of Pennsylvania

Other organizations : University of California, San Francisco

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