Invitrogen

Phospho-Histone H2A.X (Ser139) Monoclonal Antibody (3F2)

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Phospho-Histone H2A.X (Ser139) Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence staining using Product # MA1-2022, treatment with paraquat and iron induces MnSOD and Phosphorylation of H2AX in RAW 264.7 macrophages. Cells were treated for 20 hours with paraquat (500 µM) and iron (200 µg/mL) and stained with anti-Phospho-H2AX antibody.

Phospho-Histone H2A.X (Ser139) Antibody in Immunohistochemistry (IHC) — Immunohistochemistry analysis of phospho-Histone H2A.X pSer140 in postnatal mouse (PN19) lung sections. Antigen retrieval from 4% PFA, paraffin embedded sections was performed using heat induced epitope retrieval (HIER) method with sodium citrate buffer (pH 6.0). Following antigen retrieval, tissues were blocked and probed with a phospho-Histone H2A.X pSer140 mouse monoclonal antibody (Product # MA1-2022) at a dilution of 1:400. Increased staining intensity was observed in a genetic mouse model with DNA damage in airway cells.

Phospho-Histone H2A.X (Ser139) Antibody in Western Blot (WB) — Western blot analysis of Phospho-H2AX pSer140 (Product # MA1-2022) at a concentration of 1 µg/mL on Jurkat cell untreated (Lane 1) and Jurkat cell stimulated with staurosporine (Lane 2) followed by HRP conjugated goat anti-mouse lgG (H+L) Secondary Antibody.

Product Details

MA1-2022

Applications	Tested Dilution	Publications
Western Blot (WB)	1 µg/mL	View 5 publications 5 publications
Immunohistochemistry (IHC)	-	View 3 publications 3 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:400	-
Immunocytochemistry (ICC/IF)	Assay-dependent	View 8 publications 8 publications
Flow Cytometry (Flow)	1 µg / 10^6 cells	-
ELISA (ELISA)	Assay-dependent	View 1 publication 1 publication
Miscellaneous PubMed (Misc)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse
Published species	Bovine, Human, Mouse
Host/Isotype	Mouse / IgG1, kappa
Class	Monoclonal
Type	Antibody
Clone	3F2
Immunogen	Synthetic peptide sequence surrounding phosphorylated Ser140
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Protein G
Storage buffer	PBS with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_559491

Product Specific Information

MA1-2022 was used in IHC to successfully detect H2A.X pSer140 in postnatal mouse lung section.

MA1-2022 detects human and mouse phosphorylated H2AX.

MA1-2022 has been used successfully in Western blot, immunofluorescence, and ELISA procedures. By Western blot this antibody detects ~17 kDa protein representing phosphorylated H2AX in gamma irradiated HeLa cell lysate. In immunofluorescence procedures, MA1-2022 recognizes phosphorylated H2AX in gamma irradiated HeLa cells. ELISA of phosphorylated H2AX can also be performed using MA1-2022. MA1-2022 was used in IHC to successfully detect H2A.X pSer140 in postnatal mouse lung section.

The MA1-2022 immunogen is a synthetic peptide sequence surrounding phosphorylated Ser140. Some references call Ser140, by the name Ser139.

This antibody was orginally validated as part of a Thermo Scientific Cellomics High Content Screening Kit. The antibody sold separately may have slightly different performance and may need to be further optimized for the best results.

Target Information

Histone H2A.X (H2AX) is a member of the histone H2A family which is one of the four core histones making up the nucleosome core particle. In eukaryotes, DNA double strand breaks (DSBs) have been shown to trigger the phosphorylation of serine 139 at the carboxy terminus of histone H2AX resulting in gamma-H2AX. The phosphorylation of H2AX can be detected by Western blotting or immunofluorescence, revealing the frequency of DSBs. The phosphatidylinositol 3-kinases have been implicated in H2AX phosphorylation, but it is unclear if ATM is the primary H2AX kinase or if other members of the family such as DNA-PK and ATR contribute in a similar manner. Structurally, H2A.x contains 143 amino acid residues. Histone H2A.X is considered a basal histone, being synthesized in G1 as well as in S-phase, and its mRNA contains polyA addition motifs and a polyA tail along with the conserved stem-loop and U7 binding sequences involved in the processing and stability of replication type histone mRNAs. There are two forms of Histone H2A.X mRNA, one about 1600 bases long and contains polyA; the other about 575 bases long, lacking polyA. The short form behaves as a replication type histone mRNA, while the longer behaves as a basal type histone mRNA. Histone H2A.X maps to the 11q23.2-q23.3 region of the human chromosome. Histone H2A.x contributes to histone-formation and therefore the structure of DNA. Histone H2A variant H2A.x specifically regulates the interaction of MDC1 (mediator of DNA damage checkpoint protein 1), a DNA repair protein to the sites of DNA damage.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Post-translational modification of RNA m6A demethylase ALKBH5 regulates ROS-induced DNA damage response.

Nucleic acids research

Yu F,Wei J,Cui X,Yu C,Ni W,Bungert J,Wu L,He C,Qian Z

MA1-2022 was used in Immunocytochemistry to uncover a molecular mechanism involving ALKBH5 PTMs and increased mRNA m6A levels that protect genomic integrity of cells in response to ROS.

Fri Jun 04 00:00:00 EDT 2021

Regulation of meiotic progression by Sertoli-cell androgen signaling.

Molecular biology of the cell

Larose H,Kent T,Ma Q,Shami AN,Harerimana N,Li JZ,Hammoud SS,Handel MA

MA1-2022 was used in Immunohistochemistry-immunofluorescence to demonstrate that SCARKO mutant spermatocytes exhibited normal expression and localization of key protein markers of meiotic prophase events, indicating that initiation of meiotic prophase is not androgen dependent.

Tue Dec 01 00:00:00 EST 2020

Safety and bactericidal efficacy of cold atmospheric plasma generated by a flexible surface Dielectric Barrier Discharge device against Pseudomonas aeruginosa in vitro and in vivo.

Annals of clinical microbiology and antimicrobials

Dijksteel GS,Ulrich MMW,Vlig M,Sobota A,Middelkoop E,Boekema BKHL

MA1-2022 was used in Immunohistochemistry to determine the in vitro and in vivo safety and efficacy of cold atmospheric plasma generated by a surface Dielectric Barrier Discharge device.

Wed Aug 19 00:00:00 EDT 2020

"Mitotic Slippage" and Extranuclear DNA in Cancer Chemoresistance: A Focus on Telomeres.

International journal of molecular sciences

Salmina K,Bojko A,Inashkina I,Staniak K,Dudkowska M,Podlesniy P,Rumnieks F,Vainshelbaum NM,Pjanova D,Sikora E,Erenpreisa J

MA1-2022 was used in Western Blotting to explore mitotic slippage in the MDA-MB-231 cell line treated with doxorubicin.

Thu Apr 16 00:00:00 EDT 2020

Quantification of DNA Damage in Heart Tissue as a Novel Prediction Tool for Therapeutic Prognosis of Patients With Dilated Cardiomyopathy.

JACC. Basic to translational science

Ko T,Fujita K,Nomura S,Uemura Y,Yamada S,Tobita T,Katoh M,Satoh M,Ito M,Domoto Y,Hosoya Y,Amiya E,Hatano M,Morita H,Fukayama M,Aburatani H,Komuro I

MA1-2022 was used in Immunohistochemistry-immunofluorescence to develop a novel DNA-based prognosis prediction tool to assess the DNA damage in the heart tissue of patients with dilated cardiomyopathy.

Tue Oct 01 00:00:00 EDT 2019

ATM/G6PD-driven redox metabolism promotes FLT3 inhibitor resistance in acute myeloid leukemia.

Proceedings of the National Academy of Sciences of the United States of America

Gregory MA,D'Alessandro A,Alvarez-Calderon F,Kim J,Nemkov T,Adane B,Rozhok AI,Kumar A,Kumar V,Pollyea DA,Wempe MF,Jordan CT,Serkova NJ,Tan AC,Hansen KC,DeGregori J

MA12022 was used in western blot to show that inactivating ATM or G6PD sensitizes acute myeloid leukemia cells to FLT3 inhibitor-induced apoptosis

Tue Oct 25 00:00:00 EDT 2016

Elevated Expression of Hepatoma Up-Regulated Protein Inhibits γ-Irradiation-Induced Apoptosis of Prostate Cancer Cells.

Journal of cellular biochemistry

Hassan M,El Khattouti A,Ejaeidi A,Ma T,Day WA,Espinoza I,Vijayakumar S,Gomez CR

MA1-2022 was used in immunocytochemistry and western blot to characterize inhibition of gamma-irradiation-induced apoptosis of prostate cancer cells by elevated expression of hepatoma up-regulated protein

Wed Jun 01 00:00:00 EDT 2016

Elevated Expression of Hepatoma Up-Regulated Protein Inhibits γ-Irradiation-Induced Apoptosis of Prostate Cancer Cells.

Journal of cellular biochemistry

Hassan M,El Khattouti A,Ejaeidi A,Ma T,Day WA,Espinoza I,Vijayakumar S,Gomez CR

Wed Jun 01 00:00:00 EDT 2016

High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents (HPHC).

Journal of visualized experiments : JoVE

Marescotti D,Gonzalez Suarez I,Acali S,Johne S,Laurent A,Frentzel S,Hoeng J,Peitsch MC

MA1-2022 was used in immunocytochemistry to evaluate the toxicological effects of harmful and potentially harmful constituents (HPHC) by utilizing a high content screening analysis

Tue May 10 00:00:00 EDT 2016

Conservative site-specific and single-copy transgenesis in human LINE-1 elements.

Nucleic acids research

Vijaya Chandra SH,Makhija H,Peter S,Myint Wai CM,Li J,Zhu J,Ren Z,D'Alcontres MS,Siau JW,Chee S,Ghadessy FJ,Dröge P

Published figure using Phospho-Histone H2A.X (Ser140) monoclonal antibody (Product # MA1-2022) in Western Blot

Thu Apr 07 00:00:00 EDT 2016

Conservative site-specific and single-copy transgenesis in human LINE-1 elements.

Nucleic acids research

Vijaya Chandra SH,Makhija H,Peter S,Myint Wai CM,Li J,Zhu J,Ren Z,D'Alcontres MS,Siau JW,Chee S,Ghadessy FJ,Dröge P

Published figure using Phospho-Histone H2A.X (Ser140) monoclonal antibody (Product # MA1-2022) in Western Blot

Thu Apr 07 00:00:00 EDT 2016

Organization and dynamics of the nonhomologous end-joining machinery during DNA double-strand break repair.

Proceedings of the National Academy of Sciences of the United States of America

Reid DA,Keegan S,Leo-Macias A,Watanabe G,Strande NT,Chang HH,Oksuz BA,Fenyo D,Lieber MR,Ramsden DA,Rothenberg E

MA1-2022 was used in Immunofluorescence to study the nonhomolgous end-joining repair proteins present at DNA double-stranded breaks using in vivo and in vitro single-molecule methods.

Tue May 19 00:00:00 EDT 2015

A high-throughput chemical screen reveals that harmine-mediated inhibition of DYRK1A increases human pancreatic beta cell replication.

Nature medicine

Wang P,Alvarez-Perez JC,Felsenfeld DP,Liu H,Sivendran S,Bender A,Kumar A,Sanchez R,Scott DK,Garcia-Ocaña A,Stewart AF

MA1-2022 was used in immunocytochemistry to identify analogs of harmine as a new class of human beta cell mitogenic compounds and identify the pathways they activate.

Wed Apr 01 00:00:00 EDT 2015

Profiling dose-dependent activation of p53-mediated signaling pathways by chemicals with distinct mechanisms of DNA damage.

Toxicological sciences : an official journal of the Society of Toxicology

Clewell RA,Sun B,Adeleye Y,Carmichael P,Efremenko A,McMullen PD,Pendse S,Trask OJ,White A,Andersen ME

MA1-2022 was used in western blot to develop high throughput methods to assess proof-of-concept in vitro-only risk assessments

Sat Nov 01 00:00:00 EDT 2014

Comparative analysis of the cytotoxic effects of okadaic acid-group toxins on human intestinal cell lines.

Marine drugs

Ferron PJ,Hogeveen K,Fessard V,Le Hégarat L

MA1-2022 was used in immunocytochemistry to compared the toxicological effects of okadaic acid and dinophysistoxin 1 and 2

Thu Aug 21 00:00:00 EDT 2014

Comparative analysis of the cytotoxic effects of okadaic acid-group toxins on human intestinal cell lines.

Marine drugs

Ferron PJ,Hogeveen K,Fessard V,Le Hégarat L

MA1-2022 was used in immunocytochemistry to compared the toxicological effects of okadaic acid and dinophysistoxin 1 and 2

Thu Aug 21 00:00:00 EDT 2014

The NR4A2 nuclear receptor is recruited to novel nuclear foci in response to UV irradiation and participates in nucleotide excision repair.

PloS one

Jagirdar K,Yin K,Harrison M,Lim W,Muscat GE,Sturm RA,Smith AG

MA1-2022 was used in immunocytochemistry to study the mechanism underlying the role of the NR4A2 nuclear receptor in nucleotide excision repair in response to damage caused by UV irradiation

Wed Aug 20 00:00:00 EDT 2014

Cytotoxicity, fractionation and dereplication of extracts of the dinoflagellate Vulcanodinium rugosum, a producer of pinnatoxin G.

Marine drugs

Geiger M,Desanglois G,Hogeveen K,Fessard V,Leprêtre T,Mondeguer F,Guitton Y,Hervé F,Séchet V,Grovel O,Pouchus YF,Hess P

MA1-2022 was used in immunocytochemistry to isolate and identify N-carboxy-methyl-smenospongine as the likely cytotoxic and genotoxic component of extracts of Vulcanodium rugosum

Mon Sep 02 00:00:00 EDT 2013

Cytotoxicity, fractionation and dereplication of extracts of the dinoflagellate Vulcanodinium rugosum, a producer of pinnatoxin G.

Marine drugs

Geiger M,Desanglois G,Hogeveen K,Fessard V,Leprêtre T,Mondeguer F,Guitton Y,Hervé F,Séchet V,Grovel O,Pouchus YF,Hess P

MA1-2022 was used in immunocytochemistry to isolate and identify N-carboxy-methyl-smenospongine as the likely cytotoxic and genotoxic component of extracts of Vulcanodium rugosum