BioNukleo offers carefully assembled kits for the detection of (pyro-) phosphate by continuous UV-spectroscopic monitoring of 5-bromouridine phosphorolysis (PUB module, see also Eilert et
al. 2022, DOI: 10.1021/acs.analchem.1c05356). The PUB module allows quantitative phosphate detection by observing the shift in UV spectrum when
5-bromouridine and phosphate are converted to 5-bromouracil by a pyrimidine phoshphorylase. Key properties of the PUB phosphate detection system are:
Easy to use
Very few simple sample handling steps and detection by standard spectroscopic equipment.
Phosphate concentrations of 100 µM to >5 mM can readily be detected. Lower phosphate concentrations down to around 2 µM can be detected by providing 5-bromouridine in excess and
measuring absorbance differences between a baseline and a near-equilibrium state.
Highly selective to inorganic phosphate and exhibits no cross-reactivity with other phosphate species.
Compatible with different reaction volumes, reaction vessels and sizes of analysis equipment. Detection can be carried out in 1 ml
cuvettes in a photometer, 200 µl multiwell plates in a platereader or even in 1 µl droplets assessed via nanodrop.
Functioning at a broad range of experimental conditions and applicable to a variety of experimental scenarios and reactions of interest.
Compatible with parallelized experimentation, for instance in assays in multiwell plates.
No long sample incubation times and readout in under 30 mins.
Enables continuous monitoring of live reactions.
Broad temperature range
Not limited by single assay temperature requirements.
The extended version of this kit - including an inorganic pyrophosphatase (hydrolyzing pyrophosphate to orthophosphate) - enables the quantification of pyrophosphate in a fully
Available upon request. Please contact us for further information.