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MABE1177

Sigma-Aldrich

Anti-PNKP Antibody, clone H101.2

clone H101.2, from mouse

Sinónimos:

Bifunctional polynucleotide phosphatase/kinase, DNA 5′-kinase/3′-phosphatase, Polynucleotide kinase-3′-phosphatase

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

H101.2, monoclonal

species reactivity

human, bovine, monkey

technique(s)

ELISA: suitable
immunoprecipitation (IP): suitable
western blot: suitable

isotype

IgG1κ

NCBI accession no.

UniProt accession no.

shipped in

wet ice

target post-translational modification

unmodified

Gene Information

human ... PNKP(11284)

General description

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).

Specificity

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

Immunogen

Epitope: Within the N-terminal 140 amino acids.
His-tagged recombinant full-length human PNKP.

Application

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).

Quality

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.

Target description

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

Physical form

Format: Purified
Protein G 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.

Storage and Stability

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

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable


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