Purified anti-H2A.X Phospho (Ser139) Antibody

Pricing & Availability
Clone
2F3 (See other available formats)
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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Cat # Size Price Quantity Avail. Save
613401 25 µg $100
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613402 100 µg $250
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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

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 - Validated
ICFC - Reported in the literature

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

(PubMed link indicates BioLegend citation)
  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. Xiao R, et al. 2007. Mol Cell Biol. 27:5393. PubMed
  2. Mochizuki K, et al. 2008. J Cell Sci. 121:2148. PubMed
  3. Mochizuki J 2009. Mol Biol Cell. 2.519444444. PubMed
  4. Suzuki K, et al. 2010. Nucleic Acids Res. 38:e129. PubMed
  5. Massignan T, et al. 2010. J Biol Chem. 285:7752. PubMed
  6. Widodo N, et al. 2010. PLoS One. 5:e13536. PubMed
  7. Martinelli P, et al. 2011. Blood. 117:6617. PubMed
  8. Li M, et al. 2011. Mol Cell Biol. 31:2090. PubMed
  9. Hirano J 2011. J Cell Biol. 192:263. PubMed
  10. Kaul Z, et al. 2011. EMBO Rep. 13:52. PubMed
  11. Kuefner M, et al. 2012. Radiology. 264:59:00. PubMed
  12. Yoshihara Y, et al. 2012. Biochem Biophys Res Commun. 421:57. PubMed
  13. Titus S, et al. 2013. Sci Transl Med. 13:21. PubMed
  14. Jha H, et al. 2013. J Virol. 87:5255. PubMed
  15. Akbay E, et al. 2008. Am J Pathol. 173:536. PubMed
  16. Crown K, et al. 2013. g3. 3:1927. PubMed
  17. Zhdanova N, et al. 2014. Mol Cell Biol. 34:2786. PubMed
  18. Chalker S 2014. Eukaryot Cell. 13:1300. PubMed
  19. Maya-Mendoza A, et al. 2015. Mol Oncol. 9:601. PubMed
  20. Sandoval P, et al. 2015. PLoS Genet. 11: 1005405. PubMed
  21. Udugama M, et al. 2015. Nucleic Acids Res. 43: 10227 - 10237. PubMed
  22. Nyeste A, et al. 2016. J Biol Chem. 291: 4473 - 4486. PubMed
  23. Pedersen R, et al. 2016. Nat Commun. 7:13887. PubMed
  24. Feng W, et al. 2017. Proc Natl Acad Sci U S A. 114(2):406-411. PubMed
  25. Yamauchi M, et al. 2017. Sci Rep. 7:41812. PubMed
  26. Pellegrino S, et al. 2017. Cell Reports. 10.1016/j.celrep.2017.05.016. PubMed
  27. Ogata T, et al. 2017. PLoS One. 10.1371/journal.pone.0179884. PubMed
  28. Mlcochova P,et al. 2017. EMBO J.. 10.15252/embj.201796880. PubMed
  29. Valianatos G,et al. 2017. PLoS One. 10.1371/journal.pone.0185801. PubMed
  30. Yamauchi S, et al. 2017. BMC Cancer. 10.1186/s12885-017-3621-x. PubMed
  31. Massonneau J, et al. 2018. FEBS Open Bio. 8:416. PubMed
  32. Kwasna D 2018. Molecular cell. 70:150. PubMed
  33. Michelena J, et al. 2018. Nat Commun. 9:2028. PubMed
  34. Perucca P 2018. Biochimica et biophysica acta. 1865:898. PubMed
  35. Przetocka S 2018. Molecular cell. 72:568. PubMed
  36. Teloni F, et al. 2019. Mol Cell. 73:670. PubMed
  37. Zhong B, et al. 2019. Am J Cancer Res. 9:79. PubMed
  38. Taglialatela A, et al. 2017. Mol Cell. 68:414. PubMed
  39. Maffucci P, et al. 2018. J Clin Invest. 128:5489. PubMed
  40. Qin Y, et al. 2019. Mol Oncol. 13:1419. PubMed
  41. Sang M, et al. 2018. Oncol Rep. 39:2749. PubMed
  42. Paculova H, et al. 2017. Tumour Biol. 39:1010428317727479. PubMed
Publication Library
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 Phosphorylated H2A.X on UniProt.org

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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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