Protein was extracted by 4% SDS mediated lysis, followed by processing of lysates through homogenizer column (Omega). Protein concentrations were estimated by BCA (Pierce Biotechnology). Lysates were processed using PAGE with 4%–12% Bis-Tris BOLT gel (Life Technologies) and transferred to PVDF membrane (Millipore). Membranes were blocked in 10% milk/TBST and immunoblotted with the following antibodies.: rabbit monoclonal c-MYC (Cell Signaling, 1:1000), Rabbit monoclonal anti-Acetyl α Tubulin (Cell Signaling, 1:1000), Rabbit monoclonal anti-Acetylated Histone 3 (Cell Signaling, 1:1000), Rabbit polyclonal anti-Histone H3 (Cell Signaling, 1:1000), Rabbit polyclonal anti-DNMT1 (Sigma Aldrich, 1:2000), Mouse monoclonal anti-β-Actin (Sigma Aldrich, 1:10000). The loading control antibodies (anti-β-Actin, anti-Histone H3) in all cases were applied after membrane stripping.
Rabbit polyclonal anti histone h3
Manufactured by Cell Signaling Technology
Rabbit polyclonal anti-Histone H3 is a primary antibody that specifically recognizes histone H3, a core component of eukaryotic nucleosomes. It can be used to detect and quantify histone H3 in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Bolt bis tris gel, by Thermo Fisher Scientific (1 mentions)
Rabbit monoclonal anti acetyl α tubulin, by Cell Signaling Technology (1 mentions)
C myc, by Cell Signaling Technology (1 mentions)
Mouse monoclonal anti β actin, by Merck Group (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Bicinchoninic acid (bca), by Thermo Fisher Scientific (1 mentions)
Na934v, by GE Healthcare (1 mentions)
Anti cyclin d1, by Cell Signaling Technology (1 mentions)
Ecl western blotting detection reagent, by GE Healthcare (1 mentions)
2 protocols using rabbit polyclonal anti histone h3
1
Western Blot Analysis of Protein Modifications
2
Protein Expression Analysis of FOXM1/Cyclin D1/Aurora B Pathway
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Protein extractions and western blot analyses were performed as previously described (Yu et al. 2008 (link)) to determine protein expression involved in FOXM1/Cyclin D1/Aurora B pathway activation induced by Cd. The following primary antibodies were used at 1:1000 dilution over night at 4 °C: Rabbit polyclonal anti-phospho-Aurora-ABC (#2914, Cell Signaling); Rabbit polyclonal anti-total-Aurora B (#ab2254, Abcam); Mouse monoclonal anti-phospho-Histone H3ser10 (#3377, Cell signaling); Rabbit polyclonal anti-Histone H3 (#9715, Cell Signaling); Rabbit polyclonal phospho/total FOXM1 (#14655/#5436, Cell Signaling), and Rabbit polyclonal anti-cyclin D1 (#2922, Cell Signaling). Secondary antibodies used were anti-rabbit or anti-mouse IgG Horseradish peroxidase linked (NA934V, or A931V respectively, GE Healthcare, Buckingham Shire, UK). The detection kit used was the ECL Western Blotting Detection Reagent (#RPn2106, GE Healthcare). Band intensity was obtained through a Densitometer (FluorChem™, Alpha Innotech, San-Leandro, CA).
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