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Purified anti-H2A.X Phospho (Ser139) Antibody

Pricing & Availability
Clone
2F3 (See other available formats)
Regulatory Status
RUO
Other Names
H2A.x, H2a/x, Histone 2A, Histone 2A.X, Gamma-H2AX
Isotype
Mouse IgG1, κ
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Product Citations
publications
a-2F3_PURE_H2AdotX_Antibody_WB_122117
Total lysates (15 µg protein) from untreated HeLa (Lane 1) and UV treated HeLa (Lane 2) cells were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 0.1 µg/mL Purified anti-H2A.X Phospho (Ser139) antibody, clone 2F3 (upper) or 1:2000 diluted anti-GAPDH antibody, clone 1D4 (lower). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for both anti-H2A.X Phospho (Ser139) antibody and anti-GAPDH antibody. Lane M: Molecular weight ladder
  • a-2F3_PURE_H2AdotX_Antibody_WB_122117
    Total lysates (15 µg protein) from untreated HeLa (Lane 1) and UV treated HeLa (Lane 2) cells were resolved by electrophoresis (4-20% Tris-Glycine gel), transferred to nitrocellulose, and probed with 0.1 µg/mL Purified anti-H2A.X Phospho (Ser139) antibody, clone 2F3 (upper) or 1:2000 diluted anti-GAPDH antibody, clone 1D4 (lower). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for both anti-H2A.X Phospho (Ser139) antibody and anti-GAPDH antibody. Lane M: Molecular weight ladder
  • b-2F3_PURE_H2AdotX_Antibody_ICC_051019
    HeLa cells were stained with purified anti-H2A.X Phospho (Ser 139) (clone 2F3) antibody, followed by staining with DyLightTM 594 conjugated goat anti-mouse IgG (red) antibody. Actin filaments were labeled in green. Nuclei were stained with DAPI (blue).
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613401 25 µg 95€
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613402 100 µg 235€
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Description

H2A.X is a 14 kD basal histone and a member of the H2 histone family. This nuclear protein is synthesized in the G1 and S phase of the cell cycle and is known to be important for DNA repair and maintaining genomic stability and for recombination between immunoglobulin switch regions. H2A.X becomes phosphorylated on serine 139 after double-stranded DNA breaks. Phosphorylated H2A.X promotes DNA repair and maintains genomic stability. The 2F3 monoclonal antibody reacts with phosphorylated human H2A.X (Ser139) and has been shown to be useful for Western blotting, immunofluorescence and flow cytometry.

Product Details
Technical Data Sheet (pdf)

Product Details

Reactivity
Human,Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Modified peptide
Formulation
This H2A.X antibody is provided in phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide. Final antibody concentration is 0.5 mg/ml.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
Upon receipt, store between 2°C and 8°C.
Application

WB - Quality tested
ICC - Verified
ICFC - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. Western blotting, suggested working dilution(s): Use 5 µg antibody per 5 ml antibody dilution buffer for each mini-gel. For immunocytochemistry, a concentration range of 1.0 - 4.0 μg/ml is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Additional reported applications (for the relevant formats of this clone) include: immunohistochemistry on paraffin embedded sections2, immunofluorescence microscopy3-9, Western blotting 10-12, and flow cytometry1,13. Clone 2F3 cross-reacts with mouse4.

Intracellular staining protocol for Anti-H2A.X-Phosphorylated (Ser139) Antibody for Flow Cytometry

1. Prepare 70% absolute ethanol. Chill solution by storing at -20°C.
2. Prepare cells of interest.
3. Wash 1X with PBS, centrifuge at 350g for 5 min.
4. Discard the supernatant and vortex to loosen cell pellet.
5. Add pre-cooled 70% ethanol drop by drop, while vortexing.
6. Incubate at -20°C for 60 minutes.
7. Wash 3X with BioLegend Cell Staining Buffer and resuspend the cells at 0.5-1 X 107 cells/ml in the cell staining buffer.
8. Perform immunofluorescent staining for flow cytometry.

Application References
  1. Jha JC, et al. 2013. J. Virol. 87:5255. (FC) PubMed
  2. Akbay A, et al. 2008. Am J Pathol. 173:536. (IHC) PubMed
  3. Mochizuki K, et al.2008.J cell Sci.121:2148. (IF) PubMed
  4. Xiao R, et al. 2007. Mol Cell Biol.27:5393. (IF) PubMed
  5. Rossi DJ, et al. 2007. Nature. 447:725. (IF) PubMed
  6. Loidl J, et al. 2009. Mol Cell Biol. 20:2048. (IF) PubMed
  7. Beels L, et al. 2009. Circulation. 120:1903. (IF) PubMed
  8. Suzuki K, et al. 2010. Nucleic Acids Res. 38:e129. (IF) PubMed
  9. Lukaszewicz A. 2010. Chromasoma Apr 27. [Epub ahead of print] (IF) PubMed
  10. Yamada C, et al. 2010 J. Biol. Chem. 285:16693. (WB) PubMed
  11. Bu Y, et al. 2010, Biochem Biophys Res Commun. 397:157. (WB) PubMed
  12. Massignan T, et al. 2010. J. Biol Chem. 285:7752. (WB) PubMed
  13. Banath JP, et al. 2010. BMC Cancer 10:4 (FC)
  14. Zhang M., et al. 2011. Cancer Res. 23:7155. PubMed
  15. Kuefner MA, et al. 2012. Radiology 264:59. PubMed
  16. Yoshihara Y, et al. 2012. Biochem Biophys Res Cmmun. 421:57. PubMed
  17. Titus S, et al. 2013. Sci Transl Med. 13:21. PubMed
  18. Crown KN, et al. 2013. G3. 6:1927. PubMed
  19. Schenkwein D, et al. 2013. Nucleic Acids Res. 41:e61. PubMed
  20. Zhadanova NS, et al. 2014. Mol Cell Biol. 34:2786. PubMed
  21. Horrell SA, et al. 2014. Eukaryot Cell. 13:1300. PubMed
  22. Maya-Mendoza A, et al. 2015. Mol Oncol. 9:601. PubMed
Product Citations
  1. Pellegrino S, et al. 2017. Cell Reports. 10.1016/j.celrep.2017.05.016. PubMed
  2. Ogata T, et al. 2017. PLoS One. 10.1371/journal.pone.0179884. PubMed
  3. Mlcochova P,et al. 2017. EMBO J.. 10.15252/embj.201796880. PubMed
  4. Valianatos G,et al. 2017. PLoS One. 10.1371/journal.pone.0185801. PubMed
  5. Yamauchi S, et al. 2017. BMC Cancer. 10.1186/s12885-017-3621-x. PubMed
  6. Massonneau J, et al. 2018. FEBS Open Bio. 8:416. PubMed
  7. Kwasna D et al. 2018. Molecular cell. 70(1):150-164 . PubMed
  8. Michelena J, et al. 2018. Nat Commun. 9:2028. PubMed
  9. Perucca P et al. 2018. Biochimica et biophysica acta. 1865(6):898-907 . PubMed
  10. Przetocka S, et al. 2018. Mol Cell. 72:568. PubMed
  11. Teloni F et al. 2019. Mol Cell. 73(4):670-683 . PubMed
  12. Zhong B, et al. 2019. Am J Cancer Res. 9:79. PubMed
  13. Taglialatela A et al. 2017. Molecular cell. 68(2):414-430 . PubMed
  14. Maffucci P, et al. 2018. J Clin Invest. 128:5489. PubMed
  15. Qin Y, et al. 2019. Mol Oncol. 13:1419. PubMed
  16. Sang M, et al. 2018. Oncol Rep. 39:2749. PubMed
  17. Paculova H, et al. 2017. Tumour Biol. 39:1010428317727479. PubMed
  18. Saxena S et al. 2018. Cell reports. 25(12):3273-3282 . PubMed
  19. Hjorth–Jensen K, et al. 2018. Nucleic Acids Res. 46:9484. PubMed
  20. Akematsu T et al. 2017. eLife. 6 pii: e26176. PubMed
  21. Michelena J, et al. 2019. J Cell Biol. 218:2865. PubMed
  22. Delaney JR, et al. 2020. PLoS Genet. 16:e1008558. PubMed
  23. Kohutova A, et al. 2019. FASEB J. :fj201801877RR. PubMed
  24. Mlcochova P, et al. 2020. Cell Rep. 30:3972. PubMed
  25. Nguyen TM, et al. 2020. Nucleic Acids Res. 48:2621. PubMed
  26. Wang Z, et al. 2019. J Biol Chem. 294:3909. PubMed
  27. Garg J, et al. 2020. Current Biology. 29(14):2371-2379. PubMed
  28. Prendergast L, et al. 2020. Nat Commun. 3.606944444. PubMed
  29. Xu Y, et al. 2020. Oncol Rep. 44:1455. PubMed
  30. Friedman J, et al. 2018. J Immunother Cancer. 6:59. PubMed
  31. Xiao R, et al. 2007. Mol Cell Biol. 27:5393. PubMed
  32. Mochizuki K, et al. 2008. J Cell Sci. 121:2148. PubMed
  33. Mochizuki J 2009. Mol Biol Cell. 2.519444444. PubMed
  34. Suzuki K, et al. 2010. Nucleic Acids Res. 38:e129. PubMed
  35. Massignan T, et al. 2010. J Biol Chem. 285:7752. PubMed
  36. Widodo N, et al. 2010. PLoS One. 5:e13536. PubMed
  37. Martinelli P, et al. 2011. Blood. 117:6617. PubMed
  38. Li M, et al. 2011. Mol Cell Biol. 31:2090. PubMed
  39. Hirano J 2011. J Cell Biol. 192:263. PubMed
  40. Kaul Z, et al. 2011. EMBO Rep. 13:52. PubMed
  41. Kuefner M, et al. 2012. Radiology. 264:59:00. PubMed
  42. Yoshihara Y, et al. 2012. Biochem Biophys Res Commun. 421:57. PubMed
  43. Titus S, et al. 2013. Sci Transl Med. 13:21. PubMed
  44. Jha H, et al. 2013. J Virol. 87:5255. PubMed
  45. Akbay E, et al. 2008. Am J Pathol. 173:536. PubMed
  46. Crown K, et al. 2013. g3. 3:1927. PubMed
  47. Zhdanova N, et al. 2014. Mol Cell Biol. 34:2786. PubMed
  48. Chalker S 2014. Eukaryot Cell. 13:1300. PubMed
  49. Maya-Mendoza A, et al. 2015. Mol Oncol. 9:601. PubMed
  50. Sandoval P, et al. 2015. PLoS Genet. 11: 1005405. PubMed
  51. Udugama M, et al. 2015. Nucleic Acids Res. 43: 10227 - 10237. PubMed
  52. Nyeste A, et al. 2016. J Biol Chem. 291: 4473 - 4486. PubMed
  53. Pedersen R, et al. 2016. Nat Commun. 7:13887. PubMed
  54. Feng W, et al. 2017. Proc Natl Acad Sci U S A. 114(2):406-411. PubMed
  55. Yamauchi M, et al. 2017. Sci Rep. 7:41812. PubMed
  56. Gavande NS, et al. 2020. Nucleic Acids Res. 48:11536. PubMed
  57. Gatti M, et al. 2020. Cell Rep. 32:107985. PubMed
  58. Stobezki R, et al. 2020. Reprod Sci. 27:940. PubMed
  59. Hopp AK, et al. 2021. Mol Cell. 81:340. PubMed
  60. Murray-Nerger LA, et al. 2021. Nucleic Acids Res. 49:2044. PubMed
  61. Cuella-Martin R, et al. 2021. Cell. 184(4):1081-1097.e19. PubMed
  62. Swift ML, et al. 2021. Cell Reports. 34(11):108840. PubMed
  63. Sakata R, et al. 2021. Cell Reports. 35(4):108999. PubMed
  64. Stoyas CA, et al. 2020. Neuron. 630:105. PubMed
  65. Kim IK, et al. 2020. Cell Death Differ. 27:2263. PubMed
  66. Michelena J, et al. 2021. Life Sci Alliance. 4: . PubMed
  67. Jelinkova S, et al. 2021. Int J Mol Sci. 22: . PubMed
  68. Callender LA, et al. 2021. Nat Commun. 12:3379. PubMed
  69. Kychygina A, et al. 2021. Sci Rep. 11:13195. PubMed
  70. Wang R, et al. 2020. Proc Natl Acad Sci U S A. 117:33436. PubMed
  71. Suzuki T, et al. 2021. Nucleic Acids Res. 49:e40. PubMed
  72. Nomura D, et al. 2021. Biol Pharm Bull. 44:642. PubMed
  73. Roeschert I, et al. 2021. Nat Cancer. 2:312. PubMed
  74. Amrenova A, et al. 2021. PLoS One. 16:e0249059. PubMed
  75. Oshima K, et al. 2020. Nat Cancer. 1:1113. PubMed
  76. Lee S, et al. 2021. Nat Commun. 12:6309. PubMed
  77. Wang C, et al. 2020. DNA Repair (Amst). 95:102946. PubMed
  78. Switonski PM, et al. 2021. Cell Rep. 37:110062. PubMed
  79. Pelicci S, et al. 2022. Nanomaterials (Basel). 12. PubMed
  80. Gañán-Gómez I, et al. 2022. Nat Med. . PubMed
  81. Denoth-Lippuner A, et al. 2021. Cell Stem Cell. 28:2020. PubMed
  82. Kim C, et al. 2021. Nat Commun. 12:1097. PubMed
  83. Lezaja A, et al. 2021. Nat Commun. 12:3827. PubMed
  84. Ueno H, et al. 2022. Commun Biol. 5:571. PubMed
  85. Guo Y, et al. 2022. iScience. 25:104519. PubMed
  86. Schneider MWG, et al. 2022. Nature. 609:183. PubMed
  87. Furia L, et al. 2022. Int J Mol Sci. 23:. PubMed
  88. Chomiak AA, et al. 2022. iScience. 25:104354. PubMed
  89. Furia L, et al. 2022. Front Oncol. 12:960734. PubMed
  90. Xu C, et al. 2022. Cell Mol Gastroenterol Hepatol. 15:327. PubMed
  91. Hsiao HW, et al. 2023. Biomolecules. 13: . PubMed
  92. Mendoza-Munoz PL, et al. 2023. NAR Cancer. 5:zcad003. PubMed
  93. Oelschläger L, et al. 2023. Cells. 12: . PubMed
  94. Kulbay M, et al. 2022. Mol Cell Biochem. 477:2213. PubMed
  95. Marchena-Cruz E, et al. 2023. Cell Rep. 42:112148. PubMed
  96. Roggan MD, et al. 2023. Front Public Health. 11:1063250. PubMed
  97. Saez-Ayala M, et al. 2023. Nat Commun. 14:3079. PubMed
RRID
AB_315794 (BioLegend Cat. No. 613401)
AB_315795 (BioLegend Cat. No. 613402)

Antigen Details

Structure
Basal histone, H2 histone family; 14 kD
Distribution

Nuclear

Function
Phosphorylated H2AX promotes DNA repair and maintains genomic stability. Important for recombination between immunoglobulin switch regions
Modification
Phosphorylation on Ser139 after double-stranded DNA breaks
Biology Area
Cell Biology, Chromatin Remodeling/Epigenetics, DNA Repair/Replication, Neuroscience
Molecular Family
Phospho-Proteins
Antigen References

1. Mannironi C, et al.1989. Nucleic Acids Res. 17:9113.
2. Celeste A, et al. 2002. Science 296:922.
3. Bassing CH, et al. 2002. Proc. Natl. Acad. Sci. USA 99:8173.
4. Reina-San-Martin B, et al. 2003. J. Exp. Med. 197:1767.

Regulation
Synthesized in G1 and S-phase of cell cycle
Gene ID
3014 View all products for this Gene ID
UniProt
View information about H2A.X Phospho Ser139 on UniProt.org
Go To Top Version: 4    Revision Date: 11/04/2016

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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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