H2AX Antibody

Images

 
Western Blot: H2AX Antibody [NB100-383] - Samples: Whole cell lysate (15 ug) from NIH3T3 cells prepared using RIPA lysis buffer. Antibody: Affinity purified rabbit anti-H2AX antibody used for WB at 0.1 ug/ml. Detection: ...read more
Immunohistochemistry: H2AX Antibody [NB100-383] - Sample: FFPE section of human ovarian carcinoma. Antibody: Affinity purified rabbit anti-H2AX used at a dilution of 1:1,000 (1ug/ml). Detection: DAB
Western Blot: H2AX Antibody [NB100-383] - Whole cell lysate from HeLa, 293T, Jurkat, mouse TCMK-1, and mouse NIH3T3.
Western Blot: H2AX Antibody [NB100-383] - Human 293T, human G-361, and wild-type (+/+) or H2AX knockout (-/-) mouse embryonic fibroblasts. Antibody used at 0.1 ug/ml.
Western Blot: H2AX Antibody [NB100-383] - Whole cell lysate (50 ug) from 293T cells prepared using RIPA lysis buffer. Antibody: Affinity purified rabbit anti-H2AX antibody used for WB at 0.1 ug/ml. Detection: ...read more
Immunohistochemistry-Paraffin: H2AX Antibody [NB100-383] - FFPE section of human ovarian carcinoma. Antibody: Affinity purified rabbit anti-H2AX used at a dilution of 1:1,000 (1ug/ml). Detection: DAB
Proximity Ligation Assay: H2AX Antibody [NB100-383] - Secondary-conjugate Duolink II PLA in Hela cells. goat anti-human BLM (NB100-324) and rabbit anit-human H2AX (NB100-383). Image merged from DAPI (2ms) and Texas Red ...read more
Proximity Ligation Assay: H2AX Antibody [NB100-383] - Secondary-conjugate Duolink II PLA in Hela cells. goat anti-human DMAP1 (NB100-256) and rabbit anti-human H2AX (NB100-383). Image merged from DAPI (2ms) and Texas ...read more
Proximity Ligation Assay: H2AX Antibody [NB100-383] - Secondary-conjugate Duolink II PLA in Hela cells. goat anti-human ATM (NB100-271) and rabbit anti-human H2AFX (NB100-383). Image merged from DAPI (2ms) and Texas Red ...read more
Proximity Ligation Assay: H2AX Antibody [NB100-383] - Secondary-conjugate Duolink In Situ PLA in MCF-7 cells. goat anti-human POLR2A (NBP2-14859) and rabbit anti-human H2AX (NB100-383). Image merged from DAPI (2ms), ...read more
Proximity Ligation Assay: H2AX Antibody [NB100-383] - Secondary-conjugate Duolink II PLA in Hela cells. goat anti-human RAD17 (NB100-272) and rabbit anti-human H2AX (NB100-383). Image merged from DAPI (2ms) and Texas ...read more
Proximity Ligation Assay: H2AX Antibody [NB100-383] - Secondary-conjugate Duolink II PLA in Hela cells. goat anti-human SMARCA4 (NBP2-22234) and rabbit anti-human H2AX (NB100-383). Image merged from DAPI (2ms) and ...read more
Proximity Ligation Assay: H2AX Antibody [NB100-383] - Secondary-conjugate Duolink II PLA in Hela cells. goat anti-human BRD2 (NB100-260) and rabbit anti-human H2AX (NB100-383). Image merged from DAPI (2ms) and Texas ...read more
Simple Western: H2AX Antibody [NB100-383] - Simple Western lane view shows a specific band for H2AX in 0.2 mg/ml of HEK lysate(s). This experiment was performed under reducing conditions using the 12 - 230 kDa ...read more
Simple Western: H2AX Antibody [NB100-383] - Electropherogram image(s) of corresponding Simple Western lane view. H2AX antibody was used at 10 ug/ml dilution on HEK lysate(s).

Product Details

Summary
Reactivity Hu, MuSpecies Glossary
Applications WB, Simple Western, ICC/IF, IHC, IHC-P, PLA
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml
   

Biological Strategies

     

H2AX Antibody Summary

Immunogen
The immunogen recognized by this antibody maps to the C-terminus of human histone H2AX using the numbering given for Swiss-Prot entry P16104 (GeneID 3014).
Specificity
This is specific for H2AX. It is not known if this also recognizes the phosphorylated form of H2AX.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
H2AFX
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:5000-1:15000
  • Simple Western 10 ug/ml
  • Immunocytochemistry/Immunofluorescence 1:250-1:1000
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:500-1:2000
  • Proximity Ligation Assay 1:1000-1:10000
Application Notes
By Western blot, this antibody recognizes a band at ~25 kDa, representing monoubiquinated H2AX protein and ~14 kDa, representing H2AX. Co-IP application reported by Piao & co-workers (PMID: 22426396). Immunohistochemistry was reported in scientific literature.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Positive Control
gamma H2AX Lysate (NBL1-11424)
Publications
Read Publications using
NB100-383 in the following applications:

  • 1 publication
  • IHC
    1 publication
  • WB
    1 publication

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
Tris-Citrate/Phosphate (pH 7.0 - 8.0)
Preservative
0.09% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for H2AX Antibody

  • BTF antibHistone H2AX
  • Gamma H2AX
  • H2A histone family member X
  • H2A.FX
  • H2A.X
  • H2A/X
  • H2AFX
  • H2AX histone
  • H2AX
  • Hist5.2ax
  • Histone 2A
  • Histone 2A.X
  • Histone 2AX
  • Histone H2A.x
  • Histone H2A/X
  • Histone H2AFX
  • Histone H2AX
  • Including osteosarcoma
  • OSRC
  • P105 RB
  • P105RB
  • PP105
  • PP110
  • pRb
  • RB 1
  • RB1 protein
  • RB1
  • Retinoblastoma 1 (including osteosarcoma)
  • Retinoblastoma 1
  • Retinoblastoma associated protein
  • Retinoblastoma related osteosarcoma
  • Retinoblastoma susceptibility gene

Background

Histone H2AX is a 14 kDa ubiquitous member of the H2A histone family. It contains an evolutionary conserved SQ motif at the C-terminus in eukaryotes. The response of eukaryotic cells to double-strand breaks in genomic DNA includes the sequestration of many factors into nuclear foci. H2AX becomes extensively phosphorylated within 1-3 minutes of DNA damage and forms foci at break sites.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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NB100-383
Species: Hu, Mu
Applications: WB, Simple Western, ICC/IF, IHC, IHC-P, PLA

Publications for H2AX Antibody (NB100-383)(10)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 3 applications: ICC/IF, IHC, WB.


Filter By Application
ICC/IF
(1)
IHC
(1)
WB
(1)
All Applications
Filter By Species
Human
(2)
Mouse
(2)
All Species
Showing Publications 1 - 10 of 10.
Publications using NB100-383 Applications Species
Piao C, Loane DJ, Stoica BA et al. Combined inhibition of cell death induced by apoptosis inducing factor and caspases provides additive neuroprotection in experimental traumatic brain injury Neurobiology of Disease 2012 Jun [PMID: 22426396] (IHC, Mouse) IHC Mouse
Wu CY, Kang HY, Yang WL et al. Critical role of monoubiquitination of histone H2AX protein in histone H2AX phosphorylation and DNA damage response. J Biol Chem 2011 Sep [PMID: 21690091]
Artus C, Boujrad H, Bouharrour A et al. AIF promotes chromatinolysis and caspase-independent programmed necrosis by interacting with histone H2AX. EMBO J 2010 May 5 [PMID: 20360685] (Mouse) Mouse
Cheok CF, Kua N, Kaldis P, Lane DP. Combination of nutlin-3 and VX-680 selectively targets p53 mutant cells with reversible effects on cells expressing wild-type p53. Cell Death Differ 2010 Sep [PMID: 20203688] (ICC/IF, Human) ICC/IF Human
Cheng W-H, Muftic D, Muftuoglu M et al. WRN Is Required for ATM Activation the S-Phase Checkpoint in Response to Interstr Cross-Link-Induced DNA Double-Str Breaks. Mol Biol Cell;19(9):3923-3933. 2008 [PMID: 18596239]
Thompson, S L D A Compton. Examining the link between chromosomal instability aneuploidy in human cells. J Cell Biol 180: 665-672. 2008 [PMID: 18283116]
Masaki H, Nishida T, Kitajima S et al. Developmental pluripotency-associated 4 (DPPA4) localized in active chromatin inhibits mouse embryonic stem cell differentiation into a primitive ectoderm lineage. J Biol Chem 2007 Nov [PMID: 17855347]
Muramatsu Y, Ohishi T, Sakamoto M et al. Cross-species difference in telomeric function of tankyrase 1. Cancer Sci 2007 Jun [PMID: 17433040]
1 Hovest, M G, Bruggenolte, N, Hosseini, K S, Krieg, T, Herrmann, G. Senescence of Human Fibroblasts after Psoralen Photoactivation Is Mediated by ATR Kinase Persistent DNA Damage Foci at Telomeres,. 17, 1758-1767. 2006 [PMID: 16436511]
Masutomi, K et al. The telomerase reverse transcriptase regulates chromatin state DNA damage responses. PNAS 102(23): 8222-8227. 2005 [PMID: 15928077] (WB, Human) WB Human

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Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for H2AX Antibody (NB100-383). (Showing 1 - 2 of 2 FAQs).

  1. I am doing Western blot for H2AX using HL-60 cells, but I do not know which secondary antibody should I use, that is rabbit, mouse or goat?
    • Our lab recommends using a secondary that is appropriate to the primary antibody chosen for your Westerns. Therefore, if you decide to choose a Rabbit polyclonal anti-H2AX antibody, we would recommend an anti-Rabbit IgG secondary conjugated to HRP, or your specific detection system. The host of the secondary should not make a difference in your western blot assay.
  2. Could you provide the detailed immunogen peptide sequence?
    • The epitope which is recognized by NB100-383 maps to the C-terminus of human histone H2AX (Swiss-Prot entry P16104). Unfortunately we are not able to provide any further information than this as the exact immunogen information is considered to be proprietary.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

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Bioinformatics Tool for H2AX Antibody (NB100-383)

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Blogs on H2AX.

ATM - detecting and responding to DNA damage
Ataxia telangiectasia mutated (ATM) is essential for the maintenance of genomic stability. ATM is a 370 kDa serine-threonine kinase that is constitutively expressed in various tissues. Although primarily nuclear, ATM is also found at lower levels...  Read full blog post.

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Bioinformatics

Gene Symbol H2AFX
Entrez
Uniprot