For IP-Western blots, 350μg of protein from lysates was used, and MYC was pulled down with the N262 antibody (sc-764; 2μg/sample). Western blot analysis was performed as described previously [21 (link)]. Immunoblots were visualized using the Odyssey IR imager (LI-COR) that can detect both Fluor 680 and IRDye 800 secondary antibodies (1:10000). Additional antibodies used for western blot include: monoclonal pS62-MYC (1:500, Abcam), Total MYC (Y69) (1:1000, ab32072, Abcam), B56α (1:500, ab72028, Abcam), β-Actin (1:10000, A5441, Sigma) and GAPDH (1:10000, AM4300, Ambion).
Mouse tissues were collected and fixed in 10% formalin-neutral buffer. Paraffin embedding and Hematoxylin and Eosin (H&E) staining were performed at OHSU Histopathology Core. Antibodies used for IF or IHC: pS62-MYC antibody (1:100) we developed as previously described [11 (link)] and pS62-MYC (1:500, ab185656, Abcam), Ki67 antibody (1:1000, Novocastra, NCL-Ki67-MM1), anti-BrdU (1:200, MCA2060, AbD serotec), Cdk4 (1:50, sc-260, Santa Cruz Biotechnology), and CD3 (1:100, Dako). IHC images were scanned and quantified by Aperio ImageScope 11.2.0.780 (Aperio Technologies). Antibodies used for the Flow Cytometry assay was described previously [35 (link)].
Mouse tissues were collected and fixed in 10% formalin-neutral buffer. Paraffin embedding and Hematoxylin and Eosin (H&E) staining were performed at OHSU Histopathology Core. Antibodies used for IF or IHC: pS62-MYC antibody (1:100) we developed as previously described [11 (link)] and pS62-MYC (1:500, ab185656, Abcam), Ki67 antibody (1:1000, Novocastra, NCL-Ki67-MM1), anti-BrdU (1:200, MCA2060, AbD serotec), Cdk4 (1:50, sc-260, Santa Cruz Biotechnology), and CD3 (1:100, Dako). IHC images were scanned and quantified by Aperio ImageScope 11.2.0.780 (Aperio Technologies). Antibodies used for the Flow Cytometry assay was described previously [35 (link)].
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