DNA Polymerase β Assay Kit

Polymerases are the enzymes responsible for synthesizing nucleic acids. DNA Polymerase β, also known as DNA polymerase subunit beta, POLB, belongs to the Family X of DNA polymerases and functions in base excision repair (BER) . Its activity is crucial for DNA replication and drug resistance. Mitochondrial DNA is particularly prone to oxidative stress, and DNA Polymerase β is essential to maintain its stability.  DNA Polymerase β overexpression has been linked to cancer, while low levels of this enzyme can lead to hypersensitivity to alkylating agents and apoptosis. The inhibition of DNA synthesis can be used as a therapeutical approach for diseases in which cell division or energetic needs are uncontrolled, such as cancer. Inhibitors such as gemcitabine have proved efficacious in the treatment of hematological and solid tumors (including breast cancer and ovarian cancer), particularly when combined with cisplatin. The use of cisplatin results in severe side effects, so the use of DNA inhibitors can also benefit patients’ care by allowing lower doses of platinum drugs to be used.  Further studies into DNA polymerase inhibitors will bring new therapy options to cancer patients.

The DNA Polymerase β Assay Kit is a fluorogenic assay designed to measure DNA Polymerase β activity for screening and profiling applications, utilizing the fluorescent nucleic acid dye GroovyGreen™ for the quantitation of double-stranded DNA (dsDNA) products. The kit comes in a convenient 96-well format, with enough recombinant Polymerase β enzyme, DR Substrate 3, dNTP mix and buffer for 100 enzymatic reactions.Figure 1:Figure 1: DNA Polymeraseβ Assay Kit mechanism.DNA Polymerase β uses available nucleotides to synthesize a DNA strand complementary to the template strand, leading to the formation of double-stranded DNA molecules (dsDNA) . The amount of dsDNA formed directly correlates to the polymerase activity and can be quantified by the addition of a fluorescent dsDNA-binding dye which distinguishes dsDNA from ssDNA and free nucleotides. Since the dye emits fluorescence only when bound to the target dsDNA, the fluorometric readouts are low in the presence of a polymerase inhibitor.Need us to run inhibitor screens or profile your compounds against DNA Polymerase β? Check out ourDNA Replication & Repair Screening Services.

P06746

Screen molecules that inhibit polymerization activity of DNA Polymerase β in drug discovery and high throughput screening (HTS) applications.Determine compound IC50.

Catalog #NameAmountStorage21000DNA Polymerase β (POLB), His-Tag Recombinant*> 1 µg-80°CDR Substrate 336 µl-80°CdNTP Substrate Mix20 µl-80°C10x DR Buffer 3A1 ml-20°C200x GroovyGreenTMDye25 µl-20°C96-well black microplate1Room Temp* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.

-80°C

This assay kit contains DNA and will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.