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
Ave. Rating
Submit a Review
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).
See high resolution IF data...
Cat # Size Price Quantity Avail. Save
613401 25 µg $100
Check Availability


Need larger quantities of this item?
Request Bulk Quote
613402 100 µg $250
Check Availability


Need larger quantities of this item?
Request Bulk Quote
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 - 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

(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. 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
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

Related FAQs

There are no FAQs for this product.
Go To Top Version: 4    Revision Date: 11/04/2016

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

ProductsHere
Insert Note Here
Save Close Clear
Lab Timer
Tools
Login/Register
Remember me
Forgot your password? Reset Password
Request an Account