Phosphodiesterase Assay Kit (ab241034)
Key features and details
- Detection method: Fluorescent
Overview
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Product name
Phosphodiesterase Assay Kit
See all TDP2 kits -
Detection method
Fluorescent -
Product overview
The Phosphodiesterase Assay Kit (ab241034) is a fluorometric plate-based assay for kinetic measurement of total phosphodiesterase activity. It is based on the cleavage of a coumarin based synthetic substrate by phosphodiesterases to generate a product, which fluoresces at Ex/Em= 370/450 nm. It does not rely on the quantification of phosphate released by phosphodiesterase action, like other traditional methods currently in use. Therefore, sample background is negligible, and does not interfere with the assay. The substrate is specific for phosphodiesterases and is not cleaved by enzymes that act on carboxyl esters or by phosphatases. This assay can detect as low as 0.25 μU of total PDE.
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Coumarin Standard (1 mM) 1 x 100µl PDE Assay Buffer 1 x 25ml PDE Positive Control 1 vial PDE Substrate 1 x 15µl -
Research areas
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Function
DNA repair enzyme that can remove a variety of covalent adducts from DNA through hydrolysis of a 5'-phosphodiester bond, giving rise to DNA with a free 5' phosphate. Catalyzes the hydrolysis of dead-end complexes between DNA and the topoisomerase 2 (TOP2) active site tyrosine residue. Hydrolyzes 5'-phosphoglycolates on protruding 5' ends on DNA double-strand breaks (DSBs) due to DNA damage by radiation and free radicals. The 5'-tyrosyl DNA phosphodiesterase activity can enable the repair of TOP2-induced DSBs without the need for nuclease activity, creating a 'clean' DSB with 5'-phosphate termini that are ready for ligation. Has also 3'-tyrosyl DNA phosphodiesterase activity, but less efficiently and much slower than TDP1. May also act as a negative regulator of ETS1 and may inhibit nuclear factor-kappa-B activation. -
Tissue specificity
Widely expressed. -
Sequence similarities
Belongs to the CCR4/nocturin family. -
Cellular localization
Nucleus. Nucleus > PML body. - Information by UniProt
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Alternative names
- 5'-Tyr-DNA phosphodiesterase
- 5'-tyrosyl-DNA phosphodiesterase
- AD 022
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Images
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Coumarin standard curve.Coumarin standard curve.
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Phosphodiesterase reaction.Phosphodiesterase reaction in PDE (positive control), HEK 293 (40 µg protein) cells and normal human CSF (20 µL undiluted).
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Phosphodiesterase activity.Phosphodiesterase activity in HEK293 cells and CSF. Assays were performed following kit protocols.