Western blotting was carried out as previously described, using LI-COR Odyssey (LI-COR Biosciences) (Liu and Secombe 2015 (link)). Antibodies used were anti-phospho-histone H3 (#9701, 1:1000; Cell Signaling Technology), anti-histone H3 (#39763 or #39163, 1:5000; Active Motif), anti-α-tubulin (1:5000; Developmental Studies Hybridoma Bank, University of Iowa). The rabbit polyclonal KDM5 antibody was raised to amino acids 1418–1760 and has been previously published (Secombe et al. 2007 (link)).
Protocol detail
Quantitative Western Blotting Analysis
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Corresponding Organization : Albert Einstein College of Medicine
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Variable analysis
independent variables
- Phosphorylation of histone H3
- Expression of histone H3
- Expression of α-tubulin
- Expression of KDM5
- Levels of phospho-histone H3
- Levels of histone H3
- Levels of α-tubulin
- Levels of KDM5
- Western blotting protocol previously described
- Use of LI-COR Odyssey system
- Antibodies used: anti-phospho-histone H3, anti-histone H3, anti-α-tubulin, anti-KDM5
- No explicit positive controls mentioned
- No explicit negative controls mentioned
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