Chromatin Immunoprecipitation of Acetylated Histone H3
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Corresponding Organization : University of Louisville
Other organizations : Hangzhou Medical College
Variable analysis
- EpiQuik™ Tissue ChIP Kit used to perform ChIP assay
- Acetylated histone H3 antibody (1 µg) used to pre-coat the assay wells
- Normal mouse IgG (1 µl) used as a negative control to pre-coat the assay wells
- 30 mg of heart tissue cross-linked with 1% formaldehyde and the cross-link stopped by glycine solution (1.25 M)
- DNA sheared by sonication (5 pulses of 20 s each separated by 40 s rest on ice; output control: 2)
- ChIP-enriched DNA fragments precipitated, purified, and assayed by quantitative PCR
- Tissue disaggregation and nuclei isolation
- Diluted supernatants used (5 µl as input DNA, 100 µl added to the acetylated histone H3 antibody-coated wells)
- Acetylated histone H3 antibody used to pre-coat the assay wells
- Normal mouse IgG used to pre-coat the assay wells
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