FCCH025104 | FlowCellect™ Multi-Color DNA Damage Response Kit

FCCH025104
25 Tests / Kit  
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      Overview

      Replacement Information
      Description
      Catalogue NumberFCCH025104
      Trade Name
      • FlowCellect
      DescriptionFlowCellect™ Multi-Color DNA Damage Response Kit
      OverviewMillipore’s FlowCellect™ Multi-Color DNA Damage Response Kit is designed to enable a researcher a quick and easy way to detect the phosphorylation state of ATM, SMC1 and Histone H2A.X by flow cytometry. Millipore’s FlowCellect™ Multi-Color DNA Damage Response Kit was developed and tested using the DNA damaging reagent, Etoposide, in HeLa cells as a model system but the kit can be used with other human cell lines to determine the effect of mechanical and chemical reagents that can induce DNA damage through the ATM dependent pathway.
      Millipore’s FlowCellect™ Multi-Color DNA Damage Response Kit contains sufficient reagents for 25 3-color samples. The kit includes three optimized fluorescently labeled antibodies and buffers necessary for cell preparation and analysis. Detailed assay instructions are included to assist in your analysis and to ensure that the correct cell concentration is obtained during acquisition of sample data.
      Alternate Names
      • Flow Cellect
      Background InformationThe signaling pathways by which cells respond to DNA damage is an important area of study for cancer research. The mammalian cell response to DNA damage is made up of highly coordinated signaling pathways that can initiate cell cycle arrest and repair or initiate apoptosis, depending on the extent and type of the damage. Cells that are defective in DNA damage signaling pathways can cause cancer because they lack the ability to sense and repair the damage, leading to genetic instability and ultimately uncontrolled cell growth.

      The main kinase activated in response to double-stranded DNA breaks is ATM or Ataxia-telangiectasia mutated kinase. ATM is a member of the phospho inositide 3-kinase (PI3K)-related Ser/Thr protein kinase family. Inactive ATM exists as a dimer but quickly dissociates and becomes phosphorylated on Serine 1981 in response to ionizing radiation.

      Once activated, ATM phosphorylates a number of downstream factors, including P53, CHK2, SMC1, NBS1 and Histone H2A.X. SMC1 is a member of the structural maintenance of chromosome family originally identified in yeast to be important for proper segregation of the chromosomes during mitosis. However, in mammalian cells exposed to IR, phosphorylation of SMC1 on sites S957 and S966 by ATM has been shown to be required for activation of the S-phase checkpoint and is important for protection against radiation induced cell death.

      Phosphorylation of the histone variant, H2A.X, at serine 139 by ATM is an important indicator of DNA damage. As the level of DNA damage increases, the level of phospho Histone H2A.X increases and accumulates at the sites of DNA damage and is often used to indicate the level of DNA damage present within the cell (8). H2AX is also responsible for recruiting response proteins to the site of DNA damage.
      References
      Product Information
      Components
      • 1. 10X Wash Buffer: (Part No. CS202123) 1 bottle containing 13 mL
      • 2. Fixation Buffer: (Part No. CS202122) 1 bottle containing 13 mL
      • 3. 5X Assay Buffer: (Part No. CS202124) 1 bottle containing 55 mL
      • 4. 1X Permeabilization Buffer: (Part No. CS202125) 1 bottle containing 13 mL
      • 5. 20X Anti-pSMC1(S957)-AlexaFluor®488: (Part No. CS204859) 1 vial containing 150 µL
      • 6. 20X Anti-pATM(S1981)-PE: (Part No.CS204855) 1 vial containing 150 µL
      • 7. 20X Anti-pHistoneH2A.X(S139)-PerCP: (Part No. CS204852) 1 vial containing 150 µL
      Detection methodFluorescent
      Applications
      ApplicationThis FlowCellect Multi-Color DNA Damage Response Kit for Flow Cytometry is designed to enable a researcher a quick & easy way to detect the phosphorylation state of ATM, SMC1 & Histone H2A.X by flow cytometry.
      Key Applications
      • Flow Cytometry
      Biological Information
      HostMouse
      Species Reactivity
      • Human
      Antibody TypeMonoclonal Antibody
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceLot to lot in-process testing on all kit components
      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.
      • Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
      Storage and Shipping Information
      Storage Conditions4-8°C for antibodies and buffers
      Packaging Information
      Material Size25 Tests / Kit
      Transport Information
      Supplemental Information
      Specifications

      Related Products & Applications

      Software Compatibility

      Catalogue Number Description  
      0500-4120 InCyte Software for Guava® easyCyte HT Systems Show Pricing & Availability

      Product Families

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      Categories

      Life Science Research > Cell Analysis > guava easyCyte Flow Cytometers > Flow Cytometry Kits & Reagents