Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 20 mJ/cm2 ultraviolet light (UV) followed by a 30 minute recover. PVDF membrane was probed with 0.25 µg/mL of Rabbit Anti-Human Phospho-Histone H2AX (S139) Monoclonal Antibody (Catalog # MAB2288) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for Histone H2AX at approximately 17 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Phospho-Histone H2AX (S139) in Camptothecin-treated Jurkat cells by Flow Cytometry.

Naïve Jurkat cells (open histogram) or treated with 1 µM Camptothecin (1100, filled histogram) overnight were stained with Rabbit Anti-Human Phospho-Histone H2AX (S139) Monoclonal Antibody (Catalog # MAB2288), followed by PE-conjugated Anti-Rabbit IgG Secondary Antibody (F0110). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with 90% methanol. View our protocol for Staining Intracellular Molecules.

Applications for Human Phospho-Histone H2AX (S139) Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: Camptothecin-treated Jurkat cells, fixed with Flow Cytometry Fixation Buffer (Catalog# FC004) and permeabilized with 90% methanol

Western Blot

0.25 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line treated with ultraviolet light (UV)

Reviewed Applications

Read 1 review rated 5 using MAB2288 in the following applications:

Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 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.

Background: Histone H2AX

Histone H2AX is a core histone protein that 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

UniProt

P16104

Additional Histone H2AX Products

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

Product Datasheets

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COA

SDS

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

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human Phospho-Histone H2AX (S139) Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB2288

Key Product Details

Validated by

Species Reactivity

Validated:

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Label

Antibody Source

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Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

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

Detection of Human Histone H2AX by Western Blot.