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17-327 | H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection)

17-327
1 kit  192 assays
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      Overview

      Replacement Information

      Key Specifications Table

      Key ApplicationsDetection Methods
      ELISA Chemiluminescence
      Description
      Catalogue Number17-327
      Brand Family Upstate
      Trade Name
      • Upstate
      DescriptionH2A.X Phosphorylation Assay Kit (Chemiluminescence Detection)
      OverviewPhosphorylation of the histone variant H2A.X is a rapid and sensitive response to double strand DNA breaks. The H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) is a cell based assay formatted for microplate-based detection of levels of phosphorylated histone H2A.X.
      References
      Product Information
      Components
      • Anti-phospho-Histone H2A.X (Ser139), clone JBW301 (Cat.# 05-636)
      • TBS, 20X (Cat.# 20-190)
      • 20% Tween-20 (v/v)
      • 10% BSA in TBS (Blocking Buffer)
      • LumiGLO Substrate Reagent A
      • LumiGLO Substrate Reagent B
      • Goat Anti-Mouse IgG, HRP conjugate
      Detection methodChemiluminescence
      Applications
      ApplicationThe H2A.X Phosphorylation, Chemiluminescence Detection Assay Kit is a cell-based ELISA formatted for chemiluminescent detection of relative levels of phosphorylated H2A.X in microplate cells cultures.
      Key Applications
      • ELISA
      Biological Information
      Entrez Gene Number
      Entrez Gene SummaryHistones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene encodes a member of the histone H2A family, and generates two transcripts through the use of the conserved stem-loop termination motif, and the polyA addition motif.
      Gene Symbol
      • H2AFX
      • H2A/X
      • H2A.X
      • H2AX
      • H2a/x
      Modifications
      • Phosphorylation
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P16104 # Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation and for efficient repair of DNA double strand breaks (DSBs) specifically when modified by C- terminal phosphorylation.
      SIZE: 143 amino acids; 15145 Da
      SUBUNIT: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Interacts with numerous proteins required for DNA damage signaling and repair when phosphorylated on Ser-140. These include MDC1, TP53BP1, BRCA1 and the MRN complex, composed of MRE11A, RAD50, and NBN. Interaction with the MRN complex is mediated at least in part by NBN. Also interacts with DHX9/NDHII when phosphorylated on Ser-140.
      SUBCELLULAR LOCATION: Nucleus.DEVELOPMENTAL STAGE: Synthesized in G1 as well as in S-phase.
      DOMAIN: SwissProt: P16104 The [ST]-Q motif constitutes a recognition sequence for kinases from the PI3/PI4-kinase family.
      PTM: Phosphorylated on Ser-140 (to form gamma-H2AFX) in response to DNA double strand breaks (DSBs) generated by exogenous genotoxic agents and by stalled replication forks, and may also occur during meiotic recombination events and immunoglobulin class switching in lymphocytes. Phosphorylation can extend up to several thousand nucleosomes from the actual site of the DSB and may mark the surrounding chromatin for recruitment of proteins required for DNA damage signaling and repair. Widespread phosphorylation may also serve to amplify the damage signal or aid repair of persistent lesions. Phosphorylation of Ser-140 in response to ionizing radiation is mediated by both ATM and PRKDC while defects in DNA replication induce Ser-140 phosphorylation subsequent to activation of ATR and PRKDC. Dephosphorylation of Ser-140 by PP2A is required for DNA DSB repair. In meiosis, Ser-140 phosphorylation may occur at synaptonemal complexes during leptotene as an ATM-dependent response to the formation of programmed DSBs by SPO11. Ser-140 phosphorylation may subsequently occurs at unsynapsed regions of both autosomes and the XY bivalent during zygotene, downstream of ATR and BRCA1 activation. Ser-140 phosphorylation may also be required for transcriptional repression of unsynapsed chromatin and meiotic sex chromosome inactivation (MSCI), whereby the X and Y chromosomes condense in pachytene to form the heterochromatic XY-body. During immunoglobulin class switch recombination in lymphocytes, Ser-140 phosphorylation may occur at sites of DNA-recombination subsequent to activation of the activation-induced cytidine deaminase AICDA. & Monoubiquitination of Lys-120 by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression (By similarity).
      SIMILARITY: Belongs to the histone H2A family.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceThe H2A.X Phosphorylation, Chemiluminescence Detection Assay Kit is a cell-based ELISA formatted for chemiluminescent detection of relative levels of phosphorylated H2A.X in microplate cells cultures. Adherent cells are cultured in microplates, treated with agents that induce DNA damage/H2A.X phosphorylation, and are then fixed and permeabilized. Histone H2A.X phosphorylated at serine 139 is detected by Anti-phospho-H2A.X (Ser139), clone JBW301 and an anti-mouse-HRP (horseradish peroxidase) conjugate. The chemiluminescent HRP substrate LumiGLO® is then added, and signal is measured in a microplate luminometer.
      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
      Packaging Information
      Material Size1 kit
      Material Package192 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Certificates of Analysis

      TitleLot Number
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) 2469294
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - 2392252 2392252
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - 2397083 2397083
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - 2435682 2435682
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - 1993887 1993887
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - 22354A 22354A
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - 22354B 22354B
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - 25120 25120
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - 25120A 25120A
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - 30163 30163
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - DAM1567726A DAM1567726A
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - DAM1588163A DAM1588163A
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) - manual manual
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) -2510190 2510190
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) -2570151 2570151
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) -2634640 2634640
      H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) -2750304 2750304

      Brochure

      Title
      Evading Apoptosis: Cell Based Assays

      User Guides

      Title
      H2A.X Phosphorylation Assay Kit Chemiluminescence Detection

      Related Products & Applications

      Kit Component

      Catalog Number Description  
      05-636 Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301 Show Pricing & Availability
      20-190 TBS, 20X Show Pricing & Availability

      Product Families

      Categories

      Life Science Research > Cell Analysis > Cell-based Assays > Apoptosis / Cell Death Assays