Detection of Human/Mouse/Rat Histone H2AX by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line, K562 human chronic myelogenous leukemia cell line, Balb/3T3 mouse embryonic fibroblast cell line, and Nb2‑11 rat lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat Histone H2AX Monoclonal Antibody (Catalog # MAB3406) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Histone H2AX at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Histone H2AX in HeLa Human Cell Line. Histone H2AX was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat Histone H2AX Monoclonal Antibody (Catalog # MAB3406) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) 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
Reconstitute at 0.5 mg/mL in sterile PBS.
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 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.
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