General description
Chromatin Immunoprecipitation (ChIP) is a widely utilized experimental technique to monitor the association of proteins with specific DNA sequences. This technique has been used to study patterns of histone modifications and to map binding sites of various proteins on a genome-wide scale with the aid of microarray technology. ChIP has also been used to determine the temporal regulation underlying the occupation of the particular chromatin locus by multiple proteins. In addition, because formaldehyde efficiently crosslinks protein to DNA as well as protein to protein, this method can, in principle, detect proteins that bind DNA directly as well as those that bind DNA indirectly through other factors.
When performing ChIP, chromatin from cells and tissues needs to be fragmented so that it becomes soluble and resolution can be achieved in detecting protein-DNA interaction at specific loci. Sonication is a common method for producing sheared chromatin. Although this method is very efficient in releasing chromatin into the solution, it has a number of limitations. For example, foaming and overheating associated with sonication can result in protein denaturation or incomplete chromatin fragmentation. Also, sonication conditions can be physically harsh and can negatively impact immunoprecipitation efficiency. These and other limitations in the procedure can make sample sonication difficult to optimize and perform reproducibly.
The EZ-Zyme
Use:
The EZ-Zyme
Packaging
Sufficient reagents for 22 enzymatic chromatin preparations
Components
Store at 4°C:
0.5M EDTA, Catalog # 20-158. One vial containing 250 μL of 0.5M EDTA, pH 8.0.
5M NaCl, Catalog # 20-159. One vial containing 500 μL of 5M NaCl.
1M Tris-HCl, pH 6.5, Catalog # 20-160. One vial containing 500 μL of 1M Tris-HCl, pH 6.5.
10X Glycine, Catalog # 20-282. One vial containing 11 mL of 1.25M Glycine.
10X PBS, Catalog # 20-281. One vial containing 24 mL of 10X PBS.
EZ-Zyme
EZ-Zyme
EZ-Zyme
Store at -20°C:
Protease Inhibitor Cocktail II, Catalog # 20-283. One vial containing 110 μL of Protease Inhibitor Cocktail II in DMSO.
RNase A, Catalog # 20-297. One vial containing 600 μg of RNase A in 60 μL sterile water.
Proteinase K, Catalog # 20-298. One vial containing 600 μg of Proteinase K in 60 μL of 50mM Tris-HCl, pH 8.0, 10mM CaCl2.
EZ-Zyme
Storage and Stability
Upon receipt, store components at the temperatures indicated on the labels. Storage temperatures are also indicated on page 3 of this manual. Kit components are stable for 6 months from date of shipment when stored as directed.
Legal Information
EZ-Zyme is a trademark of Miltex Instrument Company, Inc.
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
- UPC:
- 41116133
- Condition:
- New
- Weight:
- 1.00 Ounces
- HazmatClass:
- No
- WeightUOM:
- LB
- MPN:
- 17-375