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APC/Fire™ 750 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
Ave. Rating
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Product Citations
publications
2F3_APCFire750_H2AXphospho_082517
Nocodazole treated HeLa cells (24 hours) were fixed and permeabilized with cold 70% ethanol, then intracellularly stained with anti-H2A.X Phospho (Ser139) (clone 2F3) APC/Fire™ 750 (filled histogram) or mouse IgG1, κ APC/Fire™ 750 (open histogram).
  • 2F3_APCFire750_H2AXphospho_082517
    Nocodazole treated HeLa cells (24 hours) were fixed and permeabilized with cold 70% ethanol, then intracellularly stained with anti-H2A.X Phospho (Ser139) (clone 2F3) APC/Fire™ 750 (filled histogram) or mouse IgG1, κ APC/Fire™ 750 (open histogram).
Compare all formats See APC/Fire™ 750 spectral data
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613421 25 tests 118€
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613422 100 tests 296€
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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

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Modified peptide
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

ICFC - Quality tested
Recommended Usage

Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood.

* APC/Fire™ 750 has a maximum excitation of 650 nm and a maximum emission of 774 nm.

Excitation Laser
Red Laser (633 nm)
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
RRID
AB_2715786 (BioLegend Cat. No. 613421)
AB_2715786 (BioLegend Cat. No. 613422)

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

Related FAQs

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Go To Top Version: 1    Revision Date: 08.28.2017

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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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.

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