PUB phosphate detection

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). 

  • 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.

 

 

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

 

 

 

Available upon request.

Please contact us for further information.