ABE2939

Anti-PrimPol

• Synonym(s): DNA-directed primase/polymerase protein, hPrimpol1;Coiled-coil domain-containing protein 111
• UNSPSC Code: 12352203
• NACRES: NA.41

Properties

• biological source: rabbit
• Quality Level: 200
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: N/A, polyclonal
• mol wt: calculated mol wt 64.41 kDa; observed mol wt ~65 kDa
• purified by: affinity chromatography
• species reactivity: human
• packaging: antibody small pack of 100 μg
• technique(s): western blot: suitable
• isotype: IgG
• epitope sequence: C-terminal
• Protein ID accession no.: NP_001332824.1
• UniProt accession no.: Q96LW4
• storage temp.: 2-8°C
• Gene Information: human ... PRIMPOL(201973)

Description

General description

DNA-directed primase/polymerase protein (UniProt: Q96LW4; also known as hPrimpol1, Coiled-coil domain-containing protein 111) is encoded by the PRIMPOL (also known as CCDC111) gene (Gene ID: 201973) in human. PrimPol is an enzyme with both DNA primase and DNA polymerase activities and is involved in DNA damage tolerance. It is detected in nuclei and cytosol with a large fraction localized inside the mitochondria. Upon UV irradiation, it is rapidly redirected to chromatin to reprime DNA synthesis. It associates with nuclear chromatin during the G1 and S phases of unperturbed cell cycles. It contains two RPA1-binding motifs (RBM) (aa 513-527 and 548-556) of which RPA1 is shown to be essential for recruitment to chromatin. It binds to the basic cleft of RPA1. The RPA2-binding motif plays a supporting role in RPA1-binding. PrimPol is required to facilitate mitochondrial and nuclear replication fork progression by initiating de novo DNA synthesis using dNTPs and acting as an error-prone DNA polymerase and is able to bypass certain DNA lesions. It displays high capacity to tolerate DNA damage lesions and is able to synthesize DNA primers downstream of lesions and allows DNA replication to continue. It is also shown to realign primers ahead of ′unreadable lesions′ thereby skipping the lesion. In vitro, PrimPol is reported to act as a DNA primase, elongating primers that PrimPol itself synthesizes, or as translesion synthesis DNA polymerase that elongates pre-existing primers across lesions. In murine models PrimPol knockout cells display hypersensitivity to DNA interstrand crosslinks-inducing drugs. Two isoforms of PrimPol have been described that are produced by alternative splicing. (Ref.: Diaz-Talavera, I., et al. (2022). Biomolecules. 12(2); 248; Gonzales-Acosta, D., et al. (2021). EMBO J. 40(14); e106355; Guilliam, TA., et al. (2017). Nat. Commun. 8:15222; Garcia-Gomez, S., et al. (2013). Mol. Cell. 52(4); 541-553; Mouron, S., et al. (2013). Nat. Struct. Mol. Biol. 20(12); 1383-1389).

Specificity

This rabbit polyclonal antibody detects DNA-directed primase/polymerase (PrimPol). It targets an epitope within 14 amino acids from the C-terminal region.

Immunogen

KLH-conjugated linear peptide corresponding to 14 amino acids from the C-terminal region of human DNA-directed primase/polymerase (PrimPol).

Application

Quality Control Applications

Evaluated by Western Blotting in U2OS cell lysate.

Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected PrimPol lysate from U2OS cells, but not in cells with PrimPol knockout.

Tested applications

Fluorescence Activated Cell Sorting (FACS): A representative lot detected PrimPol in I Fluorescence Activated Cell Sorting applications (Tirman S., et al (2021). Mol Cell. 81(19); 4026-4040).

Immunocytochemistry Analysis: A representative lot detected PrimPol in Immunocytochemistry application (Tirman S., et al (2021). Mol Cell. 81(19); 4026-4040).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Physical form

Purified rabbit polyclonal antibody in buffer containing Tris-citrate-phosphate buffer (pH 7-8) with 0.09% sodium azide

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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.

Safety Information

• Storage Class Code: 13 - Non Combustible Solids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable