Thermo Fisher Scientific Phospho-Histone H2A.X (Ser139) Monoclonal Antibody (CR55T33), eFluor 570, eBioscience

Applications and Tested Dilution

Product Specifications

Available Formats

• Unconjugated
• Alexa Fluor 488
• eFluor 660
• PE
• PerCP-eFluor 710
• Request custom conjugation

Product Specific Information

The CR55T33 monoclonal antibody recognizes phosphorylated serine 139 of human and mouse H2AX. H2AX is a member of the H2A histone family that complexes with DNA and other histones to form nucleosomes. Nucleosomes consist of approximately 147 base pairs of DNA wrapped around an octamer of histones composed of H2A, H2B, H3, and H4.

Upon DNA damage (e.g., irradiation, genotoxic stress, replication errors, or recombination), PI3K-like kinases such as ATM, ATR, and DNA-PK phosphorylate serine 139 in H2AX. This phosphorylation event is critical for recruiting DNA repair proteins.

The CR55T33 antibody detects a single band (~15 kDa) in Jurkat cells treated with etoposide.

Applications Reported:

• Immunohistochemical staining of formalin-fixed paraffin-embedded tissue sections
• Immunocytochemistry

Applications Tested:

• ICC on methanol-fixed cells ≤ 1 µg/mL
• IHC on formalin-fixed paraffin-embedded human tissue ≤ 1 µg/mL (high pH antigen retrieval recommended)
• Optimize dilution for best performance.

Filter Recommendation:
Use filters capturing 570 nm emission (e.g., Excitation 545/25, 565LP; Emission 605/70). Alexa Fluor™ 555 or TRITC filters are acceptable.
Excitation: 555 nm; Emission: 570 nm.

Target Information

Histone H2A.X (H2AX) is a member of the histone H2A family, one of four core histones forming the nucleosome. DNA double-strand breaks trigger phosphorylation of serine 139 at the C-terminus of H2AX, generating γ-H2AX, a marker for DNA damage.

Phosphorylation can be detected by Western blot or immunofluorescence. PI3K family kinases (ATM, DNA-PK, ATR) are involved in this process. H2A.X contains 143 amino acids and is synthesized in both G1 and S phases. Its mRNA exists in two forms: a 1600-base polyA form and a 575-base non-polyA form. The gene maps to chromosome 11q23.2–q23.3.

Histone H2A.X contributes to chromatin structure and mediates DNA repair protein recruitment (e.g., MDC1).

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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