H2A.X Phosphorylation Assay Kit (Flow Cytometry) (for detection of DNA Double-Strand Breaks and/or Apopto

Description
Catalogue Number	17-344
Brand Family	Upstate
Trade Name	Upstate
Description	H2A.X Phosphorylation Assay Kit (Flow Cytometry)
Overview	Phosphorylation 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.

Description

Catalogue Number

17-344

Brand Family

Upstate

Trade Name

Upstate

Description

H2A.X Phosphorylation Assay Kit (Flow Cytometry)

Overview

Phosphorylation 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.

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 method	Fluorescent

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 method

Fluorescent

Applications
Application	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.
Key Applications	Flow Cytometry

Applications

Application

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.

Key Applications

Flow Cytometry

Biological Information
Entrez Gene Number	NM_002105.2
Entrez Gene Summary	Histones 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	P16104
UniProt Summary	FUNCTION: 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.

Biological Information

Entrez Gene Number

NM_002105.2

Entrez Gene Summary

Histones 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

P16104

UniProt Summary

FUNCTION: 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.

Product Usage Statements
Quality Assurance	phosphorylation 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.

Product Usage Statements

Quality Assurance

phosphorylation 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.

Packaging Information
Material Size	1 kit
Material Package	100 assays

Packaging Information

Material Size

1 kit

Material Package

100 assays

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

H2A.X Phosphorylation Assay Kit (Flow Cytometry) (for detection of DNA Double-Strand Breaks and/or Apopto

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