xlf1 Antibody

DNA Repair Mechanisms

XLF interacts with XRCC4 to stabilize DNA bridges, facilitating repair of double-strand breaks. Phosphorylation of XLF’s disordered regions modulates its DNA-binding capacity, with phospho-mimetic mutations (e.g., Asp variants) impairing DNA bridging without affecting direct DNA binding .

Cellular Sensitivity Studies

• XLF-deficient cells exhibit hypersensitivity to DNA-damaging agents like neocarzinostatin. CRISPR/Cas9 disruption of XLF in HCT116 and CHO cells reduces DNA repair efficiency, which is partially rescued by wild-type XLF but not by phosphorylation-deficient mutants (Ala/Asp variants) .

• In XRCC4-deficient cells, XLF knockout exacerbates genomic instability, highlighting its cooperative role with XRCC4 in NHEJ .

Clinical Relevance

Mutations in XLF are linked to severe immunodeficiency, growth retardation, and increased cancer risk, underscoring the antibody’s utility in diagnosing and studying these conditions .

Table 1: XLF Antibody Performance in Key Assays

Table 2: Impact of XLF Mutations on DNA Repair

Data derived from .

Key Protocols and Validation

• Western Blotting:

  1. Use 20–30 µg of whole-cell lysate.

  2. Transfer to PVDF membrane after SDS-PAGE.

  3. Incubate with XLF antibody (1:1000) overnight at 4°C .

• Immunoprecipitation:

  • Combine 0.35 mg lysate with 1:50 antibody dilution .

  • Verify interactions with XRCC4 using secondary antibodies like VeriBlot for IP (1:5000) .

Comparative Analysis of XLF Antibodies