17-375 EZ-Zyme™ Chromatin Prep Kit

17-375
1 kit  Sufficient reagents for 22 enzymatic chromatin preparations
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      Overview

      Replacement Information

      Key Spec Table

      Description
      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.

      Use:
      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.

      uEquipment/u:
      • 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
      References
      Product Information
      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™ 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.
      Applications
      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
      Dimensions
      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
      Specifications

      Documentation

      EZ-Zyme™ Chromatin Prep Kit SDS

      Title

      Safety Data Sheet (SDS) 

      EZ-Zyme™ Chromatin Prep Kit Certificates of Analysis

      TitleLot Number
      EZ-Zyme Chromatin Prep Kit - 19593021959302
      EZ-Zyme Chromatin Prep Kit - 19924521992452
      EZ-Zyme Chromatin Prep Kit - 20222212022221
      EZ-Zyme Chromatin Prep Kit - 20322682032268
      EZ-Zyme Chromatin Prep Kit - 20645552064555
      EZ-Zyme Chromatin Prep Kit - 20879242087924
      EZ-Zyme Chromatin Prep Kit - 22120552212055
      EZ-Zyme Chromatin Prep Kit - DAM1460114DAM1460114
      EZ-Zyme Chromatin Prep Kit - DAM1514140DAM1514140
      EZ-Zyme Chromatin Prep Kit - DAM1787951DAM1787951

      References

      Reference overviewPub Med ID
      ICAM3 mediates inflammatory signaling to promote cancer cell stemness.
      Shen, W; Xie, J; Zhao, S; Du, R; Luo, X; He, H; Jiang, S; Hao, N; Chen, C; Guo, C; Liu, Y; Chen, Y; Sun, P; Yang, S; Luo, N; Xiang, R; Luo, Y
      Cancer Lett  422  29-43  2018

      Show Abstract
      29477378 29477378
      Hypoxia induces endothelial‑mesenchymal transition in pulmonary vascular remodeling.
      Zhang, B; Niu, W; Dong, HY; Liu, ML; Luo, Y; Li, ZC
      Int J Mol Med  42  270-278  2018

      Show Abstract
      29568878 29568878
      Persistent phosphorylation at specific H3 serine residues involved in chemical carcinogen-induced cell transformation.
      Zhu, X; Li, D; Zhang, Z; Zhu, W; Li, W; Zhao, J; Xing, X; He, Z; Wang, S; Wang, F; Ma, L; Bai, Q; Zeng, X; Li, J; Gao, C; Xiao, Y; Wang, Q; Chen, L; Chen, W
      Mol Carcinog  56  1449-1460  2017

      Show Abstract
      27996159 27996159
      Parathyroid Hormone-Like Hormone is a Poor Prognosis Marker of Head and Neck Cancer and Promotes Cell Growth via RUNX2 Regulation.
      Chang, WM; Lin, YF; Su, CY; Peng, HY; Chang, YC; Hsiao, JR; Chen, CL; Chang, JY; Shieh, YS; Hsiao, M; Shiah, SG
      Sci Rep  7  41131  2017

      Show Abstract
      28120940 28120940
      An Infectious Disease-Associated Il12b Polymorphism Regulates IL-12/23 p40 Transcription Involving Poly(ADP-Ribose) Polymerase 1.
      Zhao, Q; Du, Q; Wei, F; Xie, J; Ma, X
      J Immunol  198  2935-2942  2017

      Show Abstract
      28219892 28219892
      Prefoldin 1 promotes EMT and lung cancer progression by suppressing cyclin A expression.
      Wang, D; Shi, W; Tang, Y; Liu, Y; He, K; Hu, Y; Li, J; Yang, Y; Song, J
      Oncogene  36  885-898  2017

      Show Abstract
      27694898 27694898
      The transcription factor RFX5 is a transcriptional activator of the TPP1 gene in hepatocellular carcinoma.
      Zhao, Y; Xie, X; Liao, W; Zhang, H; Cao, H; Fei, R; Wang, X; Wei, L; Shao, Q; Chen, H
      Oncol Rep  37  289-296  2017

      Show Abstract
      27840983 27840983
      An hTERT/ZEB1 complex directly regulates E-cadherin to promote epithelial-to-mesenchymal transition (EMT) in colorectal cancer.
      Qin, Y; Tang, B; Hu, CJ; Xiao, YF; Xie, R; Yong, X; Wu, YY; Dong, H; Yang, SM
      Oncotarget  7  351-61  2016

      Show Abstract
      26540342 26540342
      mTORC1 regulates PTHrP to coordinate chondrocyte growth, proliferation and differentiation.
      Yan, B; Zhang, Z; Jin, D; Cai, C; Jia, C; Liu, W; Wang, T; Li, S; Zhang, H; Huang, B; Lai, P; Wang, H; Liu, A; Zeng, C; Cai, D; Jiang, Y; Bai, X
      Nat Commun  7  11151  2016

      Show Abstract
      27039827 27039827
      Inhibition of RelA expression via RNA interference induces immune tolerance in a rat keratoplasty model.
      Yang, J; Feng, S; Yi, G; Wu, W; Yi, R; Lu, X; Xu, W; Qiu, H
      Mol Immunol  73  88-97  2016

      Show Abstract
      27062711 27062711