Anti-PNKP Antibody, clone H101.2

Bifunctional polynucleotide phosphatase/kinase (EC 2.7.1.78; EC 3.1.3.32; UniProt Q96T60; also known as DNA 5′-kinase/3′-phosphatase, Polynucleotide kinase-3′-phosphatase) is encoded by the PNKP (also known as AOA4, MCSZ) gene (Gene ID 11284) in human. Polynucleotide kinase/phosphatase (PNKP) is a critical mammalian DNA repair enzyme that converts 3′-phosphate and 5′-hydroxyl at damaged DNA termini to generate 5′-phosphate and 3′-hydroxyl groups that are required for subsequent processing by DNA ligases and polymerases. PNKP dephosphorylates the 3′-phosphate termini of single nucleotide gap DNAs produced by the endonuclease VIII-like DNA glycosylases NEILs prior to gap filling and ligation during base excision repair (BER). PNKP is also involved in the the nonhomologous end joining (NHEJ) pathway-mediated double-strand breaks repair by interacting with the NHEJ scaffolding protein XRCC4. PNKP-knockdown in human cells leads to marked sensitization to spontaneous mutation and increased susceptibility to genotoxic agents. Small molecules that selectively inhibit PNKP phosphatase activity sensitize human cells to DNA damage and mutations in the PNKP phosphatase domain are associated with the developmental neurological disorder MCSZ (Microcephaly, seizures, and developmental delay).

Clone H101.2 recognizes an epitope in the N-terminal region of PNKP.

Epitope: Within the N-terminal 140 amino acids.

His-tagged recombinant full-length human PNKP.

Anti-PNKP Antibody, clone H101.2 is an antibody against PNKP for use in Western Blotting, Immunoprecipitation, ELISA.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Cell Cycle, DNA Replication & Repair

Western Blotting Analysis: 1.0 µg/mL from a representative lot detected PNKP in 50 µg of A549 whole cell lysate (Courtesy of Mesfin Fanta, University of Alberta, Canada).
Western Blotting Analysis: A representative lot detected the endogenous PNKP in A549 whole cell lysates, as well as purified full-length 60 kDa recombinant human PNKP and its 14.9 kDa N-terminal (a.a. 2-140) cyanogen bromide cleavage fragment (Fanta, M., et al. (2001). Hybridoma. 20(4):237-242).
Western Blotting Analysis: A representative lot detected the endogenous PNKP in CHO cell lysates and calf thymus tissue homogenates (Fanta, M., et al. (2001). Hybridoma. 20(4):237-242).
Immunoprecipitation Analysis: A representative lot immunoprecipited PNKP from A549 cell lysates (Fanta, M., et al. (2001). Hybridoma. 20(4):237-242).
ELISA Analysis: Clone H101.2 hybridoma culture supernatant was confirmed for its immunoreactivity against His-tagged human PNKP by ELISA (Fanta, M., et al. (2001). Hybridoma. 20(4):237-242).

Evalulated by Western Blotting in A549 cell lysate.

Western Blotting Analysis: 8.0 µg/mL of this antibody detected PNKP in 50 µg of A549 cell lysate.

~60 kDa observed. 57.08 kDa (isoform 1) and 53.00 kDa (isoform 2) calculated. Uncharacterized band(s) may appear in some lysates.

Protein G Purified

Format: Purified

Purified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Concentration: Please refer to lot specific datasheet.

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