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17-375 | EZ-Zyme™ Chromatin Prep Kit

1 kit  Sufficient reagents for 22 enzymatic chromatin preparations
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      Key Specifications Table

      Catalogue Number17-375
      Brand Family Upstate
      Trade Name
      • EZ-Zyme
      • Upstate
      DescriptionEZ-Zyme™ Chromatin Prep Kit
      OverviewChromatin 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™ Chromatin Prep kit allows ChIP analysis at nucleosome resolution by performing complete or partial digestions with a proprietary enzymatic cocktail to obtain chromatin fragments of on average one to a few nucleosomes in length. Compared to sonication, this procedure not only allows mapping the protein-DNA association at a higher resolution, but also renders the subsequent immunoprecipitation more efficient due to its mild digestion condition. In addition, enzymatic shearing is compatible with non-crosslinked ChIP (native ChIP), which allows analysis of histone modifications not only on cultured cells, but also on freshly dissected and frozen tissues.

      The EZ-Zyme™ kit contains reagents optimized for generating enzymatically processed chromatin from mammalian cells for use in chromatin immunoprecipitation. While the kit contains a full complement of buffers required for chromatin shearing, careful attention must be paid to the details of the instructions. Follow all the instructions carefully, especially with regard to incubation times and temperatures.
      Materials Required but Not DelivereduReagents/u:
      • Cells, stimulated or treated as needed for the experimental system
      • 37% Formaldehyde
      • Liquid nitrogen (N2 ) or dry ice/alcohol bath
      Note: care should be exercised in handling liquid nitrogen as liquid can damage skin or tissue on contact. Use appropriate personal protective equipment when working with liquid nitrogen or the dry ice/alcohol bath.

      • Vortex mixer
      • Timer
      • Variable volume (2-1000 μL) pipettes + tips
      • Refrigerated Microcentrifuge
      • Variable temperature water bath
      • Cell scraper
      • Microfuge tubes, 1.5 mL
      • Filter-tip pipette tips
      Product Information
      • 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™ Lysis Buffer, Catalog # 20-312. One vial containing 2.0 mL of EZ-Zyme™ Lysis Buffer.
      • EZ-Zyme™ Digestion Buffer, Catalog # 20-313. One vial containing 1.65 mL of EZ-Zyme™ Digestion Buffer.
      • EZ-Zyme™ Stop Buffer, Catalog # 20-315. One vial containing 1.65 mL of EZ-Zyme™ Stop Buffer
      • 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™ Enzymatic Cocktail, Catalog # 20-314. One vial containing 76U of Enzymatic Cocktail in 38 μL of a solution containing 50% glycerol.
      ApplicationContains proprietary reagents optimized for the enzymatic shearing of chromatin from mammalian cells at higher resolution than sonication for use in chromatin immunoprecipitation (ChIP) assays.
      Biological Information
      Physicochemical Information
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • 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.
      Storage and Shipping Information
      Storage ConditionsUpon 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.
      Packaging Information
      Material Size1 kit
      Material PackageSufficient reagents for 22 enzymatic chromatin preparations
      Transport Information
      Supplemental Information




      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      EZ-Zyme Chromatin Prep Kit - 1959302 1959302
      EZ-Zyme Chromatin Prep Kit - 1992452 1992452
      EZ-Zyme Chromatin Prep Kit - 2022221 2022221
      EZ-Zyme Chromatin Prep Kit - 2032268 2032268
      EZ-Zyme Chromatin Prep Kit - 2064555 2064555
      EZ-Zyme Chromatin Prep Kit - 2087924 2087924
      EZ-Zyme Chromatin Prep Kit - 2212055 2212055
      EZ-Zyme Chromatin Prep Kit - DAM1460114 DAM1460114
      EZ-Zyme Chromatin Prep Kit - DAM1514140 DAM1514140
      EZ-Zyme Chromatin Prep Kit - DAM1787951 DAM1787951
      EZ-Zyme Chromatin Prep Kit - DAM1828833 DAM1828833
      EZ-Zyme Chromatin Prep Kit - JBC1897223 JBC1897223
      EZ-Zymetrade; Chromatin Prep Kit - 30547 30547
      EZ-Zymetrade; Chromatin Prep Kit - JBC1369227 JBC1369227
      EZ-Zymetrade; Chromatin Prep Kit - manual manual
      EZ-Zyme™ Chromatin Prep Kit - 2504845 2504845
      EZ-Zyme™ Chromatin Prep Kit -2591320 2591320
      EZ-Zyme™ Chromatin Prep Kit -2601693 2601693
      EZ-Zyme™ Chromatin Prep Kit -2623581 2623581
      EZ-Zyme™ Chromatin Prep Kit -2642535 2642535
      EZ-Zyme™ Chromatin Prep Kit -2676614 2676614
      EZ-Zyme™ Chromatin Prep Kit -2739143 2739143
      EZ-Zyme™ Chromatin Prep Kit -2765958 2765958
      EZ-Zyme™ Chromatin Prep Kit -2820621 2820621
      EZ-Zyme™ Chromatin Prep Kit - 2382152 2382152
      EZ-Zyme™ Chromatin Prep Kit - 2391139 2391139
      EZ-Zyme™ Chromatin Prep Kit - 2325096 2325096


      Reference overviewPub Med ID
      The DEAD-box RNA helicase DDX1 interacts with RelA and enhances nuclear factor kappaB-mediated transcription.
      Musarat Ishaq,Li Ma,Xiaoyun Wu,Yongxin Mu,Ji'an Pan,Jiajie Hu,Tao Hu,Qiong Fu,Deyin Guo
      Journal of cellular biochemistry 106 2009

      Show Abstract
      19058135 19058135
      Histone modifications defining active genes persist after transcriptional and mitotic inactivation
      Kouskouti, Antigone and Talianidis, Iannis
      EMBO J, 24:347-357 (2005) 2005

      15616580 15616580
      Identification of a binding region for human origin recognition complex proteins 1 and 2 that coincides with an origin of DNA replication
      Ladenburger, Eva-Maria, et al
      Mol Cell Biol, 22:1036-1048 (2002) 2002

      11809796 11809796
      Histone H4 acetylation distinguishes coding regions of the human genome from heterochromatin in a differentiation-dependent but transcription-independent manner.
      O'Neill, L P and Turner, B M
      EMBO J., 14: 3946-57 (1995) 1995

      Show Abstract
      7664735 7664735