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