Biolegend, 613408, Alexa Fluor® 647 anti-H2A.X Phospho (Ser139) Antibody
100tests
Catalog Number: 613408
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| Verified Reactivity | Human, Mouse |
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| 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 Alexa Fluor® 647 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 immunofluorescent intracellular staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µl per million cells. * Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm. Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.View full statement regarding label licenses |
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| Excitation Laser |
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| 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. |
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| 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 |
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| Product Citations | Cao K, et al. 2022. Dev Cell. 57:1211. PubMed Georges RO, et al. 2022. Nat Commun. 13:6230. PubMed de Jong MRW, et al. 2019. Cancers (Basel). 1.66875. PubMed Kang J, et al. 2015. Proc Natl Acad Sci U S A. 112: E4236-E4245. PubMed Brel V, et al. 2011. Biochem Pharmacol. 82:1843. PubMed Tierny D, et al. 2015. Clin Cancer Res. 21: 5314 - 5323. PubMed Baba T, et al. 2021. Cell Death Dis. 12:322. PubMed Lin JR et al. 2018. eLife. 7 pii: e31657. PubMed Tamayo-Orrego L, et al. 2020. Nat Cancer. 1:840. PubMed Lin J, Sorger M 2015. Sci Rep. 6: 8390. PubMed Rivas MA, et al. 2021. Front Immunol. 12:688493. PubMed Ramkumar K, et al. 2020. Mol Cancer Res. 19:485. PubMed Yoon Y, et al. 2016. Stem Cell Reports. 7:840-853. PubMed Atashpaz S, et al. 2020. eLife. 9:e54756.. PubMed Chen R, et al. 2021. Cell Reports. 34(7):108751. PubMed Wang T et al. 2018. Immunity. 49(3):504-514 . PubMed Schenkwein D, et al. 2013. Nucleic Acids Res. 41:61. PubMed Ueno D, et al. 2022. Oncotarget. 13:1054. PubMed Staudte S, et al. 2022. Cancers (Basel). 14:. PubMed Dieter SM, et al. 2021. Cell Rep. 36:109394. PubMed Ouyang K, et al. 2009. PLoS One. 7:e10002521000252. PubMed de Jong MRW, et al. 2020. Ther Adv Hematol. 11:2040620719898373. PubMed Gu Z, et al. 2021. Nat Genet. 53:672. PubMed |
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| RRID | AB_2114994 (BioLegend Cat. No. 613407) AB_2295046 (BioLegend Cat. No. 613408) |
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| Structure | Basal histone, H2 histone family; 14 kD |
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| Distribution | Nuclear |
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| Function | Phosphorylated H2AX promotes DNA repair and maintains genomic stability. Important for recombination between immunoglobulin switch regions |
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| Modification | Phosphorylation on Ser139 after double-stranded DNA breaks |
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| Biology Area | Cell Biology, Chromatin Remodeling/Epigenetics, DNA Repair/Replication, Neuroscience |
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| Molecular Family | Phospho-Proteins |
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| 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. |
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| Regulation | Synthesized in G1 and S-phase of cell cycle |
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| Gene ID | 3014 |
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| UniProt | View information about H2A.X Phospho Ser139 on UniProt.org |
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| Clone | 2F3 |
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| Regulatory Status | RUO |
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| Other Names | H2A.x, H2a/x, Histone 2A, Histone 2A.X, Gamma-H2AX |
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| Isotype | Mouse IgG1, κ |
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