ANTI-TFAM. CLONE 20G2C12 25 Y56 (C005B-451145)

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

Clone JUAM71D is a rat monoclonal antibody that detects DNA-directed primase/polymerase (PrimPol).

Immunogen

His-tagged, full-length, recombinant human DNA-directed primase/polymerase (PrimPol).

Application

Quality Control Applications

Evaluated by Western Blotting in HeLa cell lysate.

Western Blotting Analysis: A 1:500 dilution of this antibody detected PrimPol in HeLa cell lysate.

Tested applications

Western Blotting Analysis: A representative lot detected PrimPol in Western Blotting applications (Gonzalez-Acosta, D., et al. (2021). EMBO J ;40(14); e106355).

Immunofluorescence Analysis: A representative lot detected PrimPol in Immunofluorescence applications (Gonzalez-Acosta, D., et al. (2021). EMBO J. 40(14); e106355).

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 rat monoclonal antibody IgG2b in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% 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.