Rabbit Polyclonal H2A.X antibody. Suitable for IHC-P, IP, ChIP, WB, ICC/IF and reacts with Human, Rat, Mouse samples. Cited in 30 publications.
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- Frontiers in oncology 13:11172622023The hydrophilic extract from a new tomato genotype (named DHO) kills cancer cell lines through the modulation of the DNA damage response induced by Campthotecin treatment.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDaniela Barone,Carmelina Antonella Iannuzzi,Iris Maria Forte,Maria Carmen Ragosta,Maria Cuomo,Milena Dell'Aquila,Angela Altieri,Antonella Caporaso,Rosa Camerlingo,Maria Manuela Rigano,Daria Maria Monti,Amalia Barone,Paola Imbimbo,Luigi Frusciante,Marcellino Monda,Margherita D'Angelo,Michelino De Laurentiis,Antonio Giordano,Luigi AlfanoPubMed 37409248
- Neuro-oncology advances 4:vdac1472022Targeting extracellular matrix remodeling sensitizes glioblastoma to ionizing radiation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesVarsha Thakur,Vijay S Thakur,Brittany Aguila,Tatiana I Slepak,Man Wang,Wei Song,Mohini Konai,Shahriar Mobashery,Mayland Chang,Ayush B Rana,Dazhi Wang,Juliano Tiburcio de Freitas,Sakir Humayun Gultekin,Scott M Welford,Michael E Ivan,Barbara BedogniPubMed 36212741
- Scientific reports 12:37582022Histone H2AX promotes metastatic progression by preserving glycolysis via hexokinase-2.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYue Liu,Haojian Li,Crystal N Wilson,Hui Jen Bai,Myriem Boufraqech,Urbain WeyemiPubMed 35260660
- eLife 10:2021Cavin3 released from caveolae interacts with BRCA1 to regulate the cellular stress response.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKerrie-Ann McMahon,David A Stroud,Yann Gambin,Vikas Tillu,Michele Bastiani,Emma Sierecki,Mark E Polinkovsky,Thomas E Hall,Guillermo A Gomez,Yeping Wu,Marie-Odile Parat,Nick Martel,Harriet P Lo,Kum Kum Khanna,Kirill Alexandrov,Roger Daly,Alpha Yap,Michael T Ryan,Robert G PartonPubMed 34142659
- Nature communications 12:35202021The epigenetic regulator LSH maintains fork protection and genomic stability via MacroH2A deposition and RAD51 filament formation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXiaoping Xu,Kai Ni,Yafeng He,Jianke Ren,Chongkui Sun,Yie Liu,Mirit I Aladjem,Sandra Burkett,Richard Finney,Xia Ding,Shyam K Sharan,Kathrin MueggePubMed 34112784
- NPJ aging and mechanisms of disease 7:72021Human skin aging is associated with increased expression of the histone variant H2A.J in the epidermis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesClaudia E Rübe,Caroline Bäumert,Nadine Schuler,Anna Isermann,Zoé Schmal,Matthias Glanemann,Carl Mann,Harry ScherthanPubMed 33795696
- Burns & trauma 9:tkaa0452021Pterostilbene accelerates wound healing by modulating diabetes-induced estrogen receptor β suppression in hematopoietic stem cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWeiguo Xie,Xueqing Zhou,Weigang Hu,Zhigang Chu,Qiongfang Ruan,Haimou Zhang,Min Li,Hongyu Zhang,Xiaodong Huang,Paul YaoPubMed 33654697
- Frontiers in endocrinology 11:6046482020Betulinic Acid Inhibits Endometriosis Through Suppression of Estrogen Receptor β Signaling Pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDongfang Xiang et. al.PubMed 33362719
- Cell death & disease 11:5082020Oxidized ATM promotes breast cancer stem cell enrichment through energy metabolism reprogram-mediated acetyl-CoA accumulation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDan Yang et. al.PubMed 32641713
- World journal of gastroenterology 26:2599-26172020Sleeve gastrectomy ameliorates endothelial function and prevents lung cancer by normalizing endothelin-1 axis in obese and diabetic rats.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRexiati Ruze,Ya-Cheng Xiong,Jian-Wen Li,Ming-Wei Zhong,Qian Xu,Zhi-Bo Yan,Jian-Kang Zhu,Yu-Gang Cheng,San-Yuan Hu,Guang-Yong ZhangPubMed 32523314
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA- Form
- Liquid
- Clonality
- Polyclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data
Select an application| IHC-P | IP | ChIP | WB | ICC/IF | |
|---|---|---|---|---|---|
| Human | Tested | Expected | Tested | Tested | Tested |
| Mouse | Expected | Expected | Expected | Tested | Expected |
| Rat | Expected | Tested | Expected | Expected | Expected |
IHC-P
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 0.2-0.5 µg/mL | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
IP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Rat | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
ChIP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 2 µg chromatin for 25 µg chromatin | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 1 µg/mL | Notes - |
Species Human | Dilution info 1 µg/mL | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Associated Products
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Target data
Function
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.
Alternative names
H2AFX, H2AX, Histone H2AX, H2a/x, Histone H2A.X
Recommended products
Rabbit Polyclonal H2A.X antibody. Suitable for IHC-P, IP, ChIP, WB, ICC/IF and reacts with Human, Rat, Mouse samples. Cited in 30 publications.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA- Form
- Liquid
- Clonality
- Polyclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Purification technique
- Affinity purification Immunogen
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage duration
- 1-2 weeks
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- -20°C
- Aliquoting information
- Upon delivery aliquot
- Storage information
- Avoid freeze / thaw cycle
Notes
Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.
If you do not find a host species to meet your needs, our catalogue and custom Chimeric range provides scientists the specificity of Abcam's RabMAbs in the species backbone of your choice. Remember to also review our range of edited cell lines, proteins and biochemicals relevant to your target that may help you further your research goals.
Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
8 product images
ChIP - Anti-Histone H2A.X antibody - ChIP Grade (ab20669)
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab20669 (blue), and 20μl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
Western blot - Anti-Histone H2A.X antibody - ChIP Grade (ab20669)
Lanes 1 - 3: Merged signal (red and green). Green - ab20669 observed at 17 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.ab20669 was shown to recognize Histone H2A.X in wild-type HAP1 cells as signal was lost at the expected MW in H2AFX (Histone H2A.X) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and H2AFX (Histone H2A.X) knockout samples were subjected to SDS-PAGE. ab20669 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Histone H2A.X antibody - ChIP Grade (ab20669) at 1 µg/mL
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: H2AFX (Histone H2A.X) knockout HAP1 whole cell lysate at 20 µg
Lane 3: HEK 293 whole cell lysate at 20 µg
Predicted band size: 15 kDa
Western blot - Anti-Histone H2A.X antibody - ChIP Grade (ab20669)
All lanes: Western blot - Anti-Histone H2A.X antibody - ChIP Grade (ab20669) at 1 µg/mL
Lane 1: HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg
Lane 2: HeLa Histone Preparation Nuclear Lysate at 2.5 µg
Lane 3: Histone H2A Recombinant Protein (negative control) at 0.1 µg
SecondaryAll lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 17 kDa
Exposure time: 20min
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X antibody - ChIP Grade (ab20669)
IHC image of ab20669 staining Histone H2A.X in human colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab20669, 0.5μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunoprecipitation - Anti-Histone H2A.X antibody - ChIP Grade (ab20669)
Histone H2A.X was immunoprecipitated using 0.5mg Rat Testis whole tissue extract, 5µg of Rabbit polyclonal to Histone H2A.X and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Rat Testis whole tissue extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab20669.
Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
Band: 17kDa: Histone H2A.X.All lanes: Immunoprecipitation - Anti-Histone H2A.X antibody - ChIP Grade (ab20669)
Predicted band size: 15 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X antibody - ChIP Grade (ab20669)
IHC image of Histone H2A.X staining in Human Lung formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab20669, 0.2 μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-Histone H2A.X antibody - ChIP Grade (ab20669)
All lanes: Western blot - Anti-Histone H2A.X antibody - ChIP Grade (ab20669) at 1 µg/mL
Lane 1: Lung (Mouse) Tissue Lysate at 10 µg
Lane 2: Testis (Mouse) Tissue Lysate at 10 µg
Lane 3: Lung (Rat) Tissue Lysate at 10 µg
Lane 4: Testis (Rat) Tissue Lysate - normal tissue (ab29388) at 10 µg
SecondaryAll lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 17 kDa, 55 kDa
Exposure time: 8min
Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.X antibody - ChIP Grade (ab20669)
ab20669 staining Histone H2A.X in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab20669 at 0.1µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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