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17-344 H2A.X Phosphorylation Assay Kit (Flow Cytometry)

17-344
1 kit  100 assays
Purchase on Sigma-Aldrich

Áttekintés

Replacement Information
Description
Catalogue Number17-344
Brand Family Upstate
Trade Name
  • Upstate
DescriptionH2A.X Phosphorylation Assay Kit (Flow Cytometry)
OverviewPhosphorylation of the histone variant H2A.X is a rapid and sensitive response to double strand DNA breaks. The H2A.X Phosphorylation Assay Kit (Flow cytometry) is a cell based assay formatted for flow cytometric detection of levels of phosphorylated histone H2A.X.

The H2A.X Phosphorylation Assay Kit (Flow cytometry) is a cell based assay formatted for flow cytometric detection of levels of phosphorylated Histone H2A.X. Cells are cultured in microplates, treated with agents that induce DNA damage or apoptosis, which stimulates H2A.X phosphorylation. Cells are then fixed and permeabilized in preparation for staining and detection. Histone H2A.X phosphorylated at serine 139 is detected by the addition of the anti-phospho-Histone H2A.X, FITC conjugate. Cells are then scanned in a flow cytometer to quantitate the number of cells staining positive for phosphorylated Histone H2A.X.
References
Product Information
Components
  • Anti-phospho H2A.X, FITC conjugate
  • Normal Mouse IgG, FITC conjugate (Cat.# 12-487)
  • 16X Fixation Solution
  • 10X Permeabilization Solution
  • 10X Wash Solution
Detection methodFluorescent
HS Code3002 15 90
Quality LevelMQ100
Applications
ApplicationThe H2A.X Phosphorylation Assay Kit (Flow cytometry) is a cell based assay formatted for flow cytometric detection of levels of phosphorylated Histone H2A.X.
Key Applications
  • Flow Cytometry
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 Assurancephosphorylation of Histone H2A.X
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 Package100 assays
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Katalógusszám GTIN
17-344 04053252582691

Documentation

H2A.X Phosphorylation Assay Kit (Flow Cytometry) MSDS

Title

Safety Data Sheet (SDS) 

H2A.X Phosphorylation Assay Kit (Flow Cytometry) Certificates of Analysis

TitleLot Number
H2A.X Phosphorylation Assay Kit (Flow Cytometry) 3138398
H2A.X Phosphorylation Assay Kit (Flow Cytometry) 2455658
H2A.X Phosphorylation Assay Kit (Flow Cytometry) 2955489
H2A.X Phosphorylation Assay Kit (Flow Cytometry) 2897869
H2A.X Phosphorylation Assay Kit (Flow Cytometry) 2901866
H2A.X Phosphorylation Assay Kit (Flow Cytometry) 2856124
H2A.X Phosphorylation Assay Kit (Flow Cytometry) 3077126
H2A.X Phosphorylation Assay Kit (Flow Cytometry) 2988214
H2A.X Phosphorylation Assay Kit (Flow Cytometry) 3026463
H2A.X Phosphorylation Assay Kit (Flow Cytometry) - 2119194 2119194

References

Reference overviewPub Med ID
Berberine, a genotoxic alkaloid, induces ATM-Chk1 mediated G2 arrest in prostate cancer cells.
Yu Wang,Qiao Liu,Zhaojian Liu,Boxuan Li,Zhaoliang Sun,Haibin Zhou,Xiyu Zhang,Yaoqin Gong,Changshun Shao
Mutation research  734  2011

Kivonat megmutatása
22561209 22561209
Role of progerin-induced telomere dysfunction in HGPS premature cellular senescence.
Benson, EK; Lee, SW; Aaronson, SA
J Cell Sci  123  2605-12  2009

Kivonat megmutatása Teljes cikk
20605919 20605919
The effects of G2-phase enrichment and checkpoint abrogation on low-dose hyper-radiosensitivity.
Sarah A Krueger,George D Wilson,Evano Piasentin,Michael C Joiner,Brian Marples
International journal of radiation oncology, biology, physics  77  2009

Kivonat megmutatása
20637979 20637979
BCR-ABL gene expression is required for its mutations in a novel KCL-22 cell culture model for acquired resistance of chronic myelogenous leukemia.
Hongfeng Yuan,Zhiqiang Wang,Chunggang Gao,Wengang Chen,Qin Huang,Jiing-Kuan Yee,Ravi Bhatia,WenYong Chen
The Journal of biological chemistry  285  2009

Kivonat megmutatása Teljes cikk
20007699 20007699

Brochure

Title
Advancing cancer research: From hallmarks & biomarkers to tumor microenvironment progression
Evading Apoptosis: Cell Based Assays
Shaping Epigenetics Discovery - Epigenetics Product Selection Brochure

Kapcsolódó termékek és alkalmazások

Kit Component

Katalógusszám Leírás
12-487 Normal Mouse IgG, FITC conjugate

Product Families

Kategóriák

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