📋 Workflow
Full workflow, products add-to-cart directly1
Cell lysis and histone extraction
1 hLyse cells and extract histones, optionally acid-extract to enrich
💡 Tip Add HDAC inhibitors to preserve modifications
⚠️ Caution Keep cold and fast to prevent degradation
🧪 Materials for this step (click to shop by spec/brand)
2
Quantification and separation
2 hQuantify and separate histones by SDS-PAGE or acid-urea gel
💡 Tip Small positively charged histones need special gels
⚠️ Caution Load equal amounts
🧪 Materials for this step (click to shop by spec/brand)
3
Modification-specific antibody detection
1 dayDetect modification levels by immunoblotting with modification-specific antibodies
💡 Tip Use total histone/H3 as input control
⚠️ Caution Validate antibody specificity
🧪 Materials for this step (click to shop by spec/brand)
4
ChIP and genome distribution (optional)
2–4 daysCrosslink and sonicate chromatin, immunoprecipitate with modification antibody, analyze by qPCR or sequencing
💡 Tip Include IgG negative control
⚠️ Caution Fragment chromatin uniformly
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① Uninhibited HDACs lose acetylation
- ② Unvalidated antibodies cause non-specific enrichment
- ③ Over/under sonication affects ChIP
- ④ Quantification errors give uneven loading
- ⑤ Missing input/IgG controls hamper enrichment
❓ FAQ
+How to quantify histone modifications?
Immunoblot or ChIP-qPCR/CUT&Tag for relative levels; mass spec for global stoichiometry.
+Why validate antibody specificity?
Modification antibodies cross-react; validate with peptides, mutants or cell treatments.
+ChIP vs CUT&Tag?
ChIP needs more cells and sonication; CUT&Tag needs less, is more sensitive with better signal/noise.