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Alexa Fluor® 594 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
2F3_A594_H2ApointX_Antibody_051019
HeLa cells were fixed with 1% paraformaldehyde (PFA) for ten minutes, permeabilized with 0.5% Triton X-100 for ten minutes, and blocked with 5% FBS for 30 minutes. Then the cells were intracellularly stained with 5 µg/ml of Alexa Fluor® 594 anti-H2A.X Phospho (Ser139) (clone 2F3) (red) in 5% FBS overnight followed by Alexa Fluor® 488 Phalloidin (green) staining for 20 minutes at 4°C. Nuclei were counterstained with DAPI (blue). The image was captured with 40X objective.
  • 2F3_A594_H2ApointX_Antibody_051019
    HeLa cells were fixed with 1% paraformaldehyde (PFA) for ten minutes, permeabilized with 0.5% Triton X-100 for ten minutes, and blocked with 5% FBS for 30 minutes. Then the cells were intracellularly stained with 5 µg/ml of Alexa Fluor® 594 anti-H2A.X Phospho (Ser139) (clone 2F3) (red) in 5% FBS overnight followed by Alexa Fluor® 488 Phalloidin (green) staining for 20 minutes at 4°C. Nuclei were counterstained with DAPI (blue). The image was captured with 40X objective.
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613410 100 µg 324€
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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

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.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 594 under optimal conditions.
Concentration
0.5 mg/ml
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

ICC - Quality tested
Recommended Usage

Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 1.25 - 5.0 μg/ml is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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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. Aury–Landas J, et al. 2019. Cell Physiol Biochem. 53:731. PubMed
  2. Li J, et al. 2022. J Clin Invest. Online ahead of print. PubMed
  3. Zhao B, et al. 2020. Nat Commun. 11:908. PubMed
RRID
AB_2616774 (BioLegend Cat. No. 613410)

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: 2    Revision Date: 11/04/2016

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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