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Key Documents

05-636-I

Sigma-Aldrich

Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301

clone JBW301, from mouse

Synonym(s):

Anti-phospho-Histone H2A.X (Ser139) Antibody, 05-636-I | Sigma-Aldrich

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

JBW301, monoclonal

species reactivity

human, rat, mouse

technique(s)

ChIP: suitable
immunocytochemistry: suitable
immunofluorescence: suitable
western blot: suitable

isotype

IgG1κ

NCBI accession no.

UniProt accession no.

shipped in

wet ice

target post-translational modification

phosphorylation (pSer139)

Gene Information

human ... H2AX(3014)
mouse ... H2Ax(15270)
rat ... H2Ax(500987)

General description

The histone H2A.X protein is a variant member of the H2A family of histones that is distinguished from other H2A histones by a unique carboxy-terminal sequence. This unique sequence is highly conserved throughout eukaryotic evolution and is rapidly phosphorylated by ATM or ATR at the fourth residue from the carboxy-terminus (Serine 139 in mammalian H2A.X) in response to DNA double-strand breaks (DSBs). Phosphorylation of H2A.X is important in the formation of a stable repair complex at the site of DNA damage.
H2A.X phosphorylation is a very rapid response to DNA damage, occurring within as little as one minute after exposure to ionizing radiation. Phosphorylation of H2A.X occurs irrespective of the cause of the DNA DSBs and phospho-H2A.X has been observed in response to environmental stresses that result in DSBs as well as programmed cellular events, including DNA rearrangement and apoptosis.

Immunogen

KLH-conjugated linear peptide corresponding to human phospho-Histone H2A. X (Ser139).

Application

Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301 is a highly specific mouse monoclonal antibody, that targets Histone H2A.X & has been tested in western blotting, ICC, ChIP & Immunofluorescence.
Immunocytochemistry Analysis: A 1:50-250 dilution from a representative lot detected phospho-Histone H2A. X (Ser139) in HeLa and A431 cells.
Previous lot has been demonstrated to work in Immunofluorescence and Chromatin Immunoprecipitation: See reference (Meier, Andreas et al., 2007)
Western Blotting Analysis: 0.05-1 μg/mL of this lot detected phosphorylated histone H2A.X (Ser139) in acid extracted histone lysates from Jurkat cells treated with 0.5 μM staurosporine (Catalog # 19-123).
Immunocytochemistry: 2 μg/mL of a previous lot of antibody detected phosphorylated histone H2A.X in HeLa cells treated with 0.5μM staurosporine for 4-6 hours.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Histones

Quality

Evaluated by Western Blotting in HeLa cell lysate.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected phospho-Histone H2A. X (Ser139) in 200 µg in staurosporine treated HeLa cells.

Target description

~17 kDa observed. Uncharacterized band(s) may appear in some lysates.

Physical form

Format: Purified
Protein G Purified
Purified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

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.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Kranti A Mapuskar et al.
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Ralph E Kleiner et al.
Nature chemical biology, 11(10), 807-814 (2015-09-08)
DNA double-strand break repair involves phosphorylation of histone variant H2AX ('γH2AX'), which accumulates in foci at sites of DNA damage. In current models, the recruitment of multiple DNA repair proteins to γH2AX foci depends mainly on recognition of this 'mark'

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