Biolegend, 613411, PE anti-H2A.X Phospho (Ser139) Antibody, 25tests

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.
25tests
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)
Preparation: The antibody was purified by affinity chromatography and conjugated with PE under optimal conditions.
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.
Excitation Laser: Blue Laser (488 nm)Green Laser (532 nm)/Yellow-Green Laser (561 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): 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: Otano I, et al. 2021. Nat Commun. 12:7296. PubMed Lin JR et al. 2018. eLife. 7 pii: e31657. PubMed Felker S, et al. 2022. JCI Insight. 7:. PubMed Falcone M, et al. 2022. Br J Cancer. :. PubMed Li S, et al. 2019. iScience. 16:63. PubMed
RRID: AB_2616870 (BioLegend Cat. No. 613411) AB_2616871 (BioLegend Cat. No. 613412)
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
Q: What type of PE do you use in your conjugates?
A: We use R-PE in our conjugates.