Human Phospho-Histone H2AX (S139) Antibody

R&D Systems | Catalog # AF2288

R&D Systems
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Goat

Applications

Validated:

Immunohistochemistry, Western Blot, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all Histone H2AX Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing human Histone H2AX S139 site

Specificity

Detects human Histone H2AX when phosphorylated at S139.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human Phospho-Histone H2AX (S139) Antibody

Histone H2AX antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P).

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) antibody by Western Blot.

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) antibody by Simple WesternTM.

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.
Phospho-Histone H2AX (S139) antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

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

Detection of Phospho-Histone H2AX (S139) by Western Blot

Meflufen induces DNA damage and apoptosis in cancer cells. (C) D492, D492HER2, and MDA‐MB231 cells were incubated with 1 µmol L‐1 of melflufen for 30 min and harvested on indicated timepoints and subjected for immunoblotting for H2AX/ yH2Ax. One representative blot of 3 is shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32717133), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Phospho-Histone H2AX (S139) by Western Blot

Detection of Phospho-Histone H2AX (S139) by Western Blot

Meflufen induces DNA damage and apoptosis in cancer cells. (A) D492, D492HER2, and MDA‐MB231 cells were seeded on a six well plate and incubated with either none, 1 µmol L‐1 or 5 µmol L‐1 melflufen for 30 min and harvested 3 h post‐treatment. Equally loaded protein lysates subjected for immunoblotting for H2AX/ yH2Ax One representative blot of three is shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32717133), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Phospho-Histone H2AX (S139) Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed U2OS human osteosarcoma cell line exposed to 10 Gy ionizing radiation and HeLa human cervical epithelial carcinoma cell line treated with ultraviolet radiation

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human breast cancer tissue

Simple Western

5 µg/mL
Sample: Jurkat human acute T cell leukemia cell line treated with Camptothecin (CPT)

Western Blot

0.5 µg/mL
Sample: Camptothecin-treated Jurkat human acute T cell leukemia cell line

Formulation, Preparation, and Storage

Purification

Antigen and protein A Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

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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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.

Alternate Names

H2AFX

Entrez Gene IDs

3014 (Human)

Gene Symbol

H2AX

Additional Histone H2AX Products

Product Documents for Human Phospho-Histone H2AX (S139) Antibody

Certificate of Analysis

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Product Specific Notices for Human Phospho-Histone H2AX (S139) Antibody

For research use only

Related Research Areas

Epigenetics

Citations for Human Phospho-Histone H2AX (S139) Antibody

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Protocols

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