Kits

Phosphate detection by the PUB module

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. The extended version of this kit - including an inorganic pyrophosphatase (hydrolyzing pyrophosphate to orthophosphate) - enables the quantification of pyrophosphate in a fully analogous fashion.

Key properties

Easy to use
Very few simple sample handling steps and detection by standard spectroscopic equipment.
Sensitive
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.
Selective
Highly selective to inorganic phosphate and exhibits no cross-reactivity with other phosphate species.
Scalable
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.
Versatile
Functioning at a broad range of experimental conditions and applicable to a variety of experimental scenarios and reactions of interest.
HT-suited
Compatible with parallelized experimentation, for instance in assays in multiwell plates.
Quick
No long sample incubation times and readout in under 30 mins.
Continuous
Enables continuous monitoring of live reactions.
Broad temperature range
Not limited by single assay temperature requirements.

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PUB high phosphate detection kit

Art.-No.: K-PUB-0001

PUB assay for the detection of high phosphate concentrations (>75 µM phosphate)

Kit components:

double desalted pyrimidine nucleoside phosphorylase Y02
substrate (5-bromouridine)
reaction buffer
detailed reaction protocol

for 500 reactions a 200 µl

355,00 €

Final price excl. shipping costs¹

Ships within 5-15 working days

Carefully assembled kit for the detection of high concentrations of phosphate (> 75 µM) by continuous UV-spectroscopic monitoring of 5-bromouridine phosphorolysis.

Allows for continuous and discontinuous reaction monitoring in biochemical assays to detect inorganic phosphate in solution with high accuracy and reproducibility.

Phosphate concentrations of 100 µM to >5 mM can readily be detected with standard spectroscopic 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.

For more information see also Eilert et al. 2022 (doi: 10.1021/acs.analchem.1c05356)

PUB low phosphate detection kit

Art.-No.: K-PUB-0002

PUB assay for the detection of low phosphate concentrations (5-200 µM phosphate)

Kit components:

double desalted pyrimidine nucleoside phosphorylase Y02
substrate (5-bromouridine)
reaction buffer
detailed reaction protocol

for 250 reactions a 200 µl

365,00 €

Final price excl. shipping costs¹

Ships within 5-15 working days

Carefully assembled kit for the detection of high concentrations of phosphate (> 75 µM) by continuous UV-spectroscopic monitoring of 5-bromouridine phosphorolysis.

Allows for continuous and discontinuous reaction monitoring in biochemical assays to detect inorganic phosphate in solution with high accuracy and reproducibility.

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.

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.

For more information see also Eilert et al. 2022 (doi: 10.1021/acs.analchem.1c05356)

PUB pyrophosphate detection kit

Art.-No.: K-PUB-0003

PUB assay for the detection of pyrophosphate

Kit components:

- double desalted pyrimidine nucleoside phosphorylase Y02

- substrate (5-bromouridine)

- double desalted inorganic pyrophosphatase GstIPP

- reaction buffer

- detailed reaction protocol

Available upon request

0,00 €

Final price excl. shipping costs¹

Carefully assembled kit for the detection of pyrophosphate by continuous UV-spectroscopic monitoring of 5-bromouridine phosphorolysis.

This kit is an extension of the PUB phosphate detection kit by addition of an inorganic pyrophosphatase (IPP, hydrolyzing pyrophosphate to orthophosphate) and enables the quantification of pyrophosphate in a fully analogous fashion.

Allows for continuous and discontinuous reaction monitoring in biochemical assays to detect inorganic pyrophosphate in solution with high accuracy and reproducibility.

Phosphate concentrations of 100 µM to >5 mM can readily be detected with standard spectroscopic 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.

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.

For more information see also Eilert et al. 2022 (doi: 10.1021/acs.analchem.1c05356)