The Western blot analysis was carried out as described by Zhu et al (Zhu, et al. 2014 (link)). Primary antibodies for p-AKT (#9271), total-AKT (#9272), PTEN (#9552), CDK4 (#2906), CDK6 (#3136), p-RB (#9307), MMP7 (#3801), and GAPDH (#2118) were purchased from Cell Signaling Technology (Danvers, MA). The p21 primary antibody (sc-6246), Rb (sc-50), and MMP2 (sc-10736) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Primary antibody against ERBB2 (RB-103-P0) was purchased from Neomarkers (Fremont, CA). Antibodies were used at a concentration recommended by the manufacturers. For control of protein loading, the blot was probed with the antibody against GAPDH.
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Western Blot Analysis of Cell Signaling
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Corresponding Organization : National Institutes of Health
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Variable analysis
independent variables
- Primary antibodies for p-AKT, total-AKT, PTEN, CDK4, CDK6, p-RB, MMP7, GAPDH, p21, Rb, MMP2, and ERBB2
- Protein expression levels of p-AKT, total-AKT, PTEN, CDK4, CDK6, p-RB, MMP7, GAPDH, p21, Rb, MMP2, and ERBB2
- Protein loading, as indicated by the use of GAPDH antibody
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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