Thermo Fisher Scientific Phospho-Histone H2A.X (Ser139) Monoclonal Antibody (CR55T33), PerCP-eFluor 710, eBioscience

Applications

Flow Cytometry (Flow)

• Tested Dilution: 5 µL (0.125 µg)/test
• Publications: View 2 publications

Product Specifications

Additional Formats

• Unconjugated
• Alexa Fluor 488
• eFluor 570
• eFluor 660
• PE
• 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 in eukaryotic chromatin. Following DNA damage such as double-strand breaks induced by irradiation, genotoxic stress, or replication errors, PI3K-like kinases (ATM, ATR, DNA-PK) phosphorylate serine 139 in H2AX. This phosphorylation recruits DNA repair proteins.

The antibody detects a single ~15 kDa band in reduced Jurkat cell lysates treated with etoposide.

• Applications Reported: Intracellular staining followed by flow cytometric analysis.
• Applications Tested: Pre-titrated for use with human peripheral blood cells using Foxp3/Transcription Factor Buffer Set (Product #00-5523-00).
  • Recommended use: 5 µL (0.125 µg)/test.
  • Cell count range: 10⁵–10⁸ cells/test.

Fluorochrome Information:
PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm). Use a 710/50 bandpass filter (or 695/40 as alternative). Ensure instrument compatibility.

Fixation:
Samples can be stored in IC Fixation Buffer (Product #00-8222-49) or 1-step Fix/Lyse Solution (Product #00-5333-54) for up to 3 days at 4°C in the dark. Clone-specific performance should be determined empirically.

Excitation: 488 nm
Emission: 710 nm
Laser: Blue Laser
Filtration: 0.2 µm post-manufacturing filtered

Target Information

Histone H2A.X (H2AX) is a member of the histone H2A family forming nucleosome core particles. DNA double-strand breaks (DSBs) induce phosphorylation of serine 139 at the C-terminus of H2AX, resulting in γ-H2AX, a marker for DSBs. Phosphorylation can be detected by Western blotting or immunofluorescence.

H2AX phosphorylation is mediated by phosphatidylinositol 3-kinases such as ATM, DNA-PK, and ATR. H2A.X contains 143 amino acids and is synthesized in both G1 and S phases. Two mRNA forms exist: a 1600-base polyA+ form and a 575-base replication-type form. Human H2A.X maps to chromosome 11q23.2–q23.3 and plays a key role in chromatin structure and DNA repair protein recruitment.

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

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