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DNA Polymerase θ Assay Kit

CAT: 0421-82107Size: 96 ReactionsDry Ice: YesHazardous: No
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CAT#:0421-82107Size:96 Reactions
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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.
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 Polymeraseθ 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 ourDNA Replication & Repair Screening Services.
UniProt
O75417
Applications
Screen molecules that inhibit polymerization activity of polymerase θ in drug discovery and high throughput screening (HTS) applications.Determine compound IC50.
Format
Catalog #NameAmountStorage101945DNA Polymerase θ, His-Tag, SUMO-Tag Recombinant*>1 µg-80°C82775DR Substrate 336 µl-80°C82776dNTP Substrate Mix20 µl-80°C8277710x DR Buffer 3A2 x 1 ml-80°C82778200x GroovyGreenTMDye25 µl-80°C7968596-well black microplate1Room Temp* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.
Shipping Conditions
-80°C
Storage Conditions
This assay kit contains DNA and will perform optimally for up to6 monthsfrom date of receipt when the materials are stored as directed.
Notes
Troubleshooting GuideVisitbpsbioscience.com/assay-kits-faqfor detailed troubleshooting instructions. For all further questions, please email[email protected]
Contraindications
The final concentration of DMSO in the assay should not exceed 1%.Fluorescent compounds that have λex=502 nm and detection at λem=523 nm can interfere with the readouts.It is recommended that the compound alone is tested to determine any potential interference of the compound on the assay results.

UniProtKB · O75417

DNA polymerase theta

DPOLQ_HUMAN · Homo sapiens

View on UniProt ↗
Primary accession
O75417
Review status
UniProtKB reviewed (Swiss-Prot)
Gene
POLQ
Protein existence
1: Evidence at protein level
Organism
Homo sapiens (Human)
Taxonomy ID
9606
Alternative names
—
EC number
—
Processing
—
Secondary accessions
O95160, Q6VMB5
Protein keywords

Technical term

3D-structureProteomics identificationReference proteome

PTM

AcetylationPhosphoprotein

Coding sequence diversity

Alternative splicing

Ligand

ATP-bindingNucleotide-binding

Cellular component

ChromosomeNucleus

Biological process

DNA damageDNA repair

Molecular function

DNA-directed DNA polymeraseHelicaseHydrolaseIsomeraseMultifunctional enzymeNucleotidyltransferaseTransferase

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