Biolegend, 613410, Alexa Fluor® 594 anti-H2A.X Phospho (Ser139) Antibody, 100μg

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.
100μg
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.View full statement regarding label licenses
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): Jha JC, et al. 2013. J. Virol. 87:5255. (FC) PubMed Akbay A, et al. 2008. Am J Pathol. 173:536. (IHC) PubMed Mochizuki K, et al.2008.J cell Sci.121:2148. (IF) PubMed Xiao R, et al. 2007. Mol Cell Biol.27:5393. (IF) PubMed Rossi DJ, et al. 2007. Nature. 447:725. (IF) PubMed Loidl J, et al. 2009. Mol Cell Biol. 20:2048. (IF) PubMed Beels L, et al. 2009. Circulation. 120:1903. (IF) PubMed Suzuki K, et al. 2010. Nucleic Acids Res. 38:e129. (IF) PubMed Lukaszewicz A. 2010. Chromasoma Apr 27. [Epub ahead of print] (IF) PubMed Yamada C, et al. 2010 J. Biol. Chem. 285:16693. (WB) PubMed Bu Y, et al. 2010, Biochem Biophys Res Commun. 397:157. (WB) PubMed Massignan T, et al. 2010. J. Biol Chem. 285:7752. (WB) PubMed Banath JP, et al. 2010. BMC Cancer 10:4 (FC) Zhang M., et al. 2011. Cancer Res. 23:7155. PubMed Kuefner MA, et al. 2012. Radiology 264:59. PubMed Yoshihara Y, et al. 2012. Biochem Biophys Res Cmmun. 421:57. PubMed Titus S, et al. 2013. Sci Transl Med. 13:21. PubMed Crown KN, et al. 2013. G3. 6:1927. PubMed Schenkwein D, et al. 2013. Nucleic Acids Res. 41:e61. PubMed Zhadanova NS, et al. 2014. Mol Cell Biol. 34:2786. PubMed Horrell SA, et al. 2014. Eukaryot Cell. 13:1300. PubMed Maya-Mendoza A, et al. 2015. Mol Oncol. 9:601. PubMed
Product Citations: Aury–Landas J, et al. 2019. Cell Physiol Biochem. 53:731. PubMed Li J, et al. 2022. J Clin Invest. Online ahead of print. PubMed Zhao B, et al. 2020. Nat Commun. 11:908. PubMed
RRID: AB_2616774 (BioLegend Cat. No. 613410)
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
UniProt: View information about H2A.X Phospho Ser139 on UniProt.org
Clone: 2F3
Regulatory Status: RUO
Other Names: H2A.x, H2a/x, Histone 2A, Histone 2A.X, Gamma-H2AX
Isotype: Mouse IgG1, κ