Biological Information

Background Information:

DNA Polymerase I (or Pol I) is an enzyme that participates in the process of DNA replication. Pol I possesses DNA-dependent DNA polymerase, exonuclease and RNA-dependent DNA polymerase proofreading activities. Most DNA polymerases participate in specialized DNA repair and DNA damage tolerance processes. The activity of DNA polymerases may affect the response of a cell to DNA-damaging carcinogens and chemotherapeutic agents. The inhibition of DNA polymerase activities could be useful as an adjuvant to DNA-damaging therapies and for chemotherapeutic benefit.

Family:

DNA Repair & Mitotic Regulation

Sub Family:

DNA Polymerase

Protein Name:

DNA Polymerase Alpha

Uniprot Number:

P09884

Gene Name:

POLA1

Gene ID:

5422

Gene Aliases:

p180

Accession Number:

NC_000913

Organism:

Bacteria

Construct Details:

Large (Klenow) Fragment, Recombinant

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Antagonist

Detection Method:

Radiometry

Testing Information

Procedure Summary:

DNA polymerase I from E. coli is used. Test compound or vehicle and two oligonucleotides primer 27AA and template 171AA are preincubated with 0.15 U/ml‡ of enzyme for 15 minutes at 37⁰C in Tris buffer pH 8.0. The reaction is initiated by addition of 150 nM unlabeled TTP and 150 nM [methyl-3H] TTP for another 15 minute incubation period. The reaction is terminated by adding EDTA and SPA. Read the plate in a Top Count Packard counter. Compounds are screened at 10 μM.

Substrate:

0.3 µM TTP + [³H]TTP

Incubation:

15 min at 37°C

Control Inhibitor:

Rifampicin

Criteria For Significance:

≥ 50 of max stimulation or inhibition

Test Sample Requirements:

Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).

Minimum Order Quantity:

2

Turnaround Time

Standard:

15 Business Days

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

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