DNA Polymerase θ Assay Kit

Catalog #

82107

$825 *

Size: 96 reactions

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Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

The DNA Polymerase θ Assay Kit is a fluorogenic assay designed to measure DNA Polymerase θ 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 and contains enough recombinant DNA Polymerase θ enzyme (amino acids 1792-2590), DR Substrate 3, dNTP mix and buffer for 100 enzymatic reactions.

Figure 1: DNA Polymerase θ Assay Kit mechanism.
DNA Polymerase θ 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 θ? Check out our DNA Replication & Repair Screening Services.

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Synonyms

DNA Polymerase Theta, POLQ

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Product Data Gallery

Inhibition of Pol β, γ and θ polymerase activity by nervonic acid and activity in the presence of G-quadruplex DNA variant.

Product Info

Storage and Usage

Citations

Assay Kit Format

Fluorogenic

Materials Required But Not Supplied

Fluorescence plate reader capable of measurement at λex502/λem523 nm.

Format

Catalog #	Name	Amount	Storage
101945	DNA Polymerase θ, His-Tag, SUMO-Tag Recombinant*	>1 µg	-80°C
	DR Substrate 3	1 µg	-80°C
	dNTP Substrate Mix	20 µl	-80°C
	10x DR Buffer 3A	2 x 1 ml	-80°C
	200x GroovyGreen^TM Dye	25 µl	-80°C
	96-well black microplate	1	Room Temp

* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.

Background

Polymerases are the enzymes responsible for synthesizing nucleic acids. DNA Polymerase θ, also known as DNA polymerase subunit theta, POLQ, belongs to the Family A of DNA polymerases. In addition to its DNA polymerase activity, it acts on the theta-mediated end joining (TMEJ), which classifies it as a specialized polymerase. TMEJ is a DNA repair mode that is prone to errors, and in addition to DNA polymerase θ it involves PARP1 (poly polymerase 1) and DNA ligase III. Inhibition of DNA polymerase θ combined with a homologous recombination (HR) deficiency, as in BRCA (breast cancer type 1/2 susceptibility protein) deficient cells, is synthetic lethal. This makes its inhibitors candidates for HR deficient cancers. It can also enhance the effect of PARP inhibitors and be used in cases where resistance to PARP inhibitors has occurred. Further development of DNA polymerase θ inhibitors will bring new therapy options to cancer patients, potentially bypassing the dose-limiting toxicities seen when using PARP inhibitors combined with conventional chemotherapy.

References

Berdis A., 2017, Front. Mol. Biosci. 4:  https://doi.org/10.3389/fmolb.2017.00078

Zatreanu D., et al., 2021 Nature Communications 12: 3636.