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Ultra Chromatin Immunoprecipitation Kit
Table of Contents
CHP2 Kit Components and Storage ...........................................................................................................2
Product Description..............................................................................................................................................3
CHP2 is Designed for ChIP-Seq......................................................................................................................3
Staph A DNA Contamination is Reduced Using ChIP Next Gen Seq Sepharose................4
Workflow.....................................................................................................................................................................4
Bridging Antibody Requirement, and Stability......................................................................................5
Reagents and Equipment Recommended But Not Provided.......................................................5
Preparation Instructions.....................................................................................................................................6
Table 1: Guidelines for Cell Number Requirements...................................................................6
Critical Parameters for ChIP-Seq.....................................................................................................................7
Experimental Design............................................................................................................................................7
Procedure...................................................................................................................................................................8
Section 1. Blocking ChIP Next Gen Seq Sepharose....................................................................8
Section 2. Preparation of Cross-linked Cells...............................................................................8-9
Section 3. Preparation of Cross-linked Chromatin...............................................................9-10
Section 4. Pre-clearing of Cross-linked Chromatin.................................................................. 11
Section 5. Antibody Binding................................................................................................................ 11
Section 6. Bridging Antibody and IP................................................................................................ 12
Section 7. Washings.................................................................................................................................. 13
Section 8. ChIP Elution............................................................................................................................ 13
Section 9. Cross-link Reversal............................................................................................................... 13
Section 10. DNA Purification................................................................................................................ 14
Section 11. Q-PCR Assays...................................................................................................................... 14
Section 12. Data Analysis....................................................................................................................... 15
Section 13. Expected Results............................................................................................................... 16
Section 14. Preparation for ChIP-chip............................................................................................. 16
Section 15. Preparation for ChIP-Seq .............................................................................................. 16
References .............................................................................................................................................................. 16
Troubleshooting Guide..............................................................................................................................17-18
Appendix................................................................................................................................................................. 19
Chromatin Sonication Optimization.................................................................................19
Cross-link Reversal and DNA Purification.................................................................19-20
Application Data (Figs. 3-9)......................................................................................................20-24
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