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

The 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.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Key Applications: ELISA
17-327
1 kit  192 assays
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

Key ApplicationsDetection Methods
ELISAChemiluminescence
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
Quality LevelMQ100
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
Global Trade Item Number
Catalogue Number GTIN
17-327 04053252010859

Documentation

H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) Certificates of Analysis

TitleLot Number
H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) 2879062
H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) 2984503
H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) 2906030
H2A.X Phosphorylation Assay Kit (Chemiluminescence Detection) 2899016
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
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Title
H2A.X Phosphorylation Assay Kit Chemiluminescence Detection

Related Products & Applications

Kit Component

Catalogue Number Description
05-636 Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301
20-190 TBS, 20X

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