Imprint® Chromatin Immunoprecipitation Kit
SIGMA/CHP1 - Complete ChIP reaction in 6 hours in flexible strip well format
Synonym: Chromatin Immunoprecipitation
Product Type: Chemical
ChIP workflow
Comparison of time required for protocol completion from fixation through purification using different Chromatin Immunoprecipitation kits. Each ChIP experiment was carried out using each manufacturers recommended protocol and total required times were compared.
One day is sufficient to acquire ChIP enriched DNA using Sigma′s Imprint ChIP protocol. ChIP with antibodies against mouse IgG, H3K9ac, RNAP and H3K27me3 was completed on 10,000 and 100,000 SW480 cells per reaction. A fraction (1%) of the sonicated chromatin was set aside as ′input′ DNA before the antibody affinity manipulations. DNA was purified following crosslink reversal. The resulting enriched DNA was probed for GAPDH using specific primers by qPCR. Percent input was calculated by 100 × 2^(Ctadjusted Input – CtEnriched). Input DNA Ct was adjusted from 1% to 100% equivalent by subtracting 6.644 Cts or Log2 100.
Figure 3. A highly sensitive ChIP assay. HeLa cells were counted, fixed, and immunoprecipitated according to the Imprint CHP1 bulletin with optional High Sensitivity method. ChIP assays were performed using Anti-H3K9ac (H9286) and kit provided antibodies against Human RNA Polymerase II and non-specific Mouse IgG. To evaluate ChIP enrichment, SYBR qPCR was conducted targeting the GAPDH promoter region (highly expressed house-keeping gene). Percent Input describes the amount of DNA with and without antibody selection. For these antibodies, apparent enrichment increased with fewer cells per reaction from diminished non-specific pull-down.
Figure 4. A highly sensitive ChIP assay. HeLa cells were counted, fixed, and immunoprecipitated according to the Imprint CHP1 bulletin with optional High Sensitivity method. ChIP assays were performed using Anti-H3K9ac (H9286) and kit provided antibodies against Human RNA Polymerase II and non-specific Mouse IgG. To evaluate ChIP enrichment, SYBR qPCR was conducted targeting the GAPDH promoter region (highly expressed house-keeping gene). Percent Input describes the amount of DNA with and without antibody selection. For these antibodies, apparent enrichment increased with fewer cells per reaction from diminished non-specific pull-down.
| Quality Level | 200  |
| storage temp. | −20°C |
| Application: | The Imprint Chromatin Immunoprecipitation Kit is a flexible, rapid and complete method for investigating protein-DNA interactions by chromatin immunoprecipitation (ChIP). • Suitable for downstream applications • Individual target characterization to genome-wide profiling techniques • Characterization of signal transduction pathways • Verification of ChIp-chIP and ChIP-seq data |
| Features and Benefits: | • Fast—Total protocol time of less than 6 hours making the Imprint kit the fastest on the market • Sensitive—As few as 10,000 cells required for each ChIP sample • Convenient—Fewest steps of any available ChIP protocol • Flexible—Protocols for cells or tissue, and convenient strip-well format for high-throughput applications • Complete—Includes columns and reagents for DNA purification as well as an integrated protocol for amplification with the GenomePlex technology |
| General description: | Imprint Chromatin Immunoprecipitation Kit provides a complete solution for Chromatin Immunoprecipitation including columns and reagents for DNA purification and an integrated protocol for ChIP DNA amplification with our GenomePlex® Whole Genome Amplification Kit (WGA2) . The flexible format allows for immunoprecipitation and purification of DNA from mammalian cells or tissue in a convenient strip-well format. For Frequently Asked Questions about this kit, please see ChIP Troubleshooting Questions . |
| Legal Information: | GenomePlex is a registered trademark of Takara Bio USA, Inc. |
| Legal Information: | Imprint is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Symbol |     GHS02,GHS05,GHS07,GHS08 |
| Signal word | Danger |
| Hazard statements | H225 - H290 - H302 - H315 - H319 - H334 - H336 - H412 |
| Precautionary statements | P210 - P233 - P273 - P301 + P312 - P303 + P361 + P353 - P305 + P351 + P338 |
| Hazard Codes | F,Xn |
| Risk Statements | 11-22-36/38-42-67 |
| Safety Statements | 9-16-23-26-33-45 |
| RIDADR | UN 3316 9 |
| WGK Germany | WGK 3 |
| Flash Point(F) | 53.6 °F - closed cup |
| Flash Point(C) | 12 °C - closed cup |
| Storage Temp. | −20°C |
| UNSPSC | 41116158 |
| Components | Column Preparation Solution; Proteinase K from Tritirachium album, buffered aqueous glycerol solution, Molecular Biology, ≥800 units/mL; Protease Inhibitor Cocktail, for use with mammalian cell and tissue extracts, DMSO solution; Normal Mouse IgG antibody produced in mouse; Anti-RNA polymerase II from mouse |