Human Phospho-Histone H2AX (S139) Antibody

Catalog # Availability Size / Price Qty
AF2288
AF2288-SP
Histone H2AX in Human Breast Cancer Tissue.
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Product Details
Citations (8)
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Human Phospho-Histone H2AX (S139) Antibody Summary

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. View Larger

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.

Western Blot Detection of Human Phospho-Histone H2AX (S139) by Western Blot. View Larger

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.

Simple Western Detection of Human Phospho-Histone H2AX (S139) by Simple WesternTM. View Larger

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. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

Product Datasheets

Citations for Human Phospho-Histone H2AX (S139) Antibody

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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  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 Types: Whole Tissue
    Applications: 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 Types: Cell Lysates
    Applications: 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 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 Types: Cell Lysates
    Applications: WB
  5. 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 Types: Cell Lysates
    Applications: WB
  6. 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 Types: Whole Cells
    Applications: ICC
  7. 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 Types: Whole Cells
    Applications: 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 Types: Whole Cells
    Applications: ICC

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