Human Phospho-Histone H2AX (S139) Antibody

(8 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Histone H2AX when phosphorylated at S139.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen and protein A Affinity-purified
  • Immunogen
    Phosphopeptide containing human Histone H2AX S139 site
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Simple Western
    5 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunohistochemistry

Histone H2AX in Human Breast Cancer Tissue. Histone H2AX was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human Phospho-Histone H2AX (S139) by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 μM camptothecin (CPT) for for the indicated times. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Histone H2AX (S139) at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Phospho-Histone H2AX (S139) by Simple WesternTM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 µM Camptothecin (CPT) for 24 hours, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Histone H2AX (S139) at approximately 25 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Immunocytochemistry

Phospho-Histone H2AX (S139) in HeLa Human Cell Line. Histone H2AX phosphorylated at S139 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with ultraviolet radiation using Rabbit Anti-Human Phospho-Histone H2AX (S139) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2288) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Histone H2AX
Histone H2AX is phosphorylated at S139 in cells exposed to DNA double-strand break-inducing agents, such as ionizing radiation. The S139 phosphorylated H2AX, termed ( gamma -H2AX, marks the site of DNA double-strand breaks and serves to recruit cell cycle checkpoint and DNA repair factors to the site of damage.
  • Entrez Gene IDs:
    3014 (Human)
  • Alternate Names:
    H2AFX; Histone H2AX
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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Species
Applications
Sample Type
  1. SMN deficiency in severe models of spinal muscular atrophy causes widespread intron retention and DNA damage
    Authors: M Jangi, C Fleet, P Cullen, SV Gupta, S Mekhoubad, E Chiao, N Allaire, CF Bennett, F Rigo, AR Krainer, JA Hurt, JP Carulli, JF Staropoli
    Proc. Natl. Acad. Sci. U.S.A, 2017;0(0):.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC
  2. Synergistic Activity of Deguelin and Fludarabine in Cells from Chronic Lymphocytic Leukemia Patients and in the New Zealand Black Murine Model
    Authors: N Rebolleda, I Losada-Fer, G Perez-Chac, R Castejon, S Rosado, M Morado, MT Vallejo-Cr, A Martinez, JA Vargas-Nuñ, P Perez-Acie
    PLoS ONE, 2016;11(4):e0154159.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Cyclin-dependent kinase five mediates activation of lung xanthine oxidoreductase in response to hypoxia.
    Authors: Kim, Bo S, Serebreni, Leonid, Fallica, Jonathan, Hamdan, Omar, Wang, Lan, Johnston, Laura, Kolb, Todd, Damarla, Mahendra, Damico, Rachel, Hassoun, Paul M
    PLoS ONE, 2015;10(4):e0124189.
  4. Inhibition of topoisomerase II by 8-chloro-adenosine triphosphate induces DNA double-stranded breaks in 8-chloro-adenosine-exposed human myelocytic leukemia K562 cells.
    Authors: Yang SY, Jia XZ, Feng LY, Li SY, An GS, Ni JH, Jia HT
    Biochem. Pharmacol., 2009;77(3):433-43.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  5. Inhibition of CHK1 kinase by Go6976 converts 8-chloro-adenosine-induced G2/M arrest into S arrest in human myelocytic leukemia K562 cells.
    Authors: Jia XZ, Yang SY, Zhou J, Li SY, Ni JH, An GS, Jia HT
    Biochem. Pharmacol., 2009;77(5):770-80.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  6. Induction of MHC class I-related chain B (MICB) by 5-aza-2'-deoxycytidine.
    Authors: Tang KF, He CX, Zeng GL, Wu J, Song GB, Shi YS, Zhang WG, Huang AL, Steinle A, Ren H
    Biochem. Biophys. Res. Commun., 2008;370(4):578-83.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  7. DNA cross-linking, double-strand breaks, and apoptosis in corneal endothelial cells after a single exposure to mitomycin C.
    Authors: Roh DS, Cook AL, Rhee SS, Joshi A, Kowalski R, Dhaliwal DK, Funderburgh JL
    Invest. Ophthalmol. Vis. Sci., 2008;49(11):4837-43.
    Species: Goat
    Sample Type: Whole Cells
    Application: ICC
  8. Histone H2AX and Fanconi anemia FANCD2 function in the same pathway to maintain chromosome stability.
    Authors: Bogliolo M, Lyakhovich A, Callen E, Castella M, Cappelli E, Ramirez MJ, Creus A, Marcos R, Kalb R, Neveling K, Schindler D, Surralles J
    EMBO J., 2007;26(5):1340-51.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
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