Chromatin Immunoprecipitation Assay Protocol
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Corresponding Organization : La Jolla Institute For Allergy & Immunology
Other organizations : Indiana University – Purdue University Indianapolis, Johnson & Johnson (United States), Janssen (United States)
Variable analysis
- Serum starvation of FLS for 24 hours
- Formaldehyde fixation (1%) for 8 minutes at room temperature
- Antibody used for chromatin immunoprecipitation (anti-H3K4me1, anti-GR, and rabbit IgG)
- Enrichment of DNA regions immunoprecipitated with the specified antibodies, measured by qPCR
- Sonication of chromatin
- Incubation of chromatin-antibody complexes overnight at 4°C
- Immunoprecipitation of complexes with protein A magnetic beads for 1 hour at 4°C
- Reversal of cross-linking at 65°C for 2 hours
- DNA purification
- 10% input sample
- Rabbit IgG
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