PRECICE® PRPP-S Assay Kit

Spectrophotometric, 96‑well microplate format

Phosphoribosylpyrophosphate synthetase (P; EC 2.7.6.1) is an essential enzyme in the purine salvage pathway, encoded by the PRPS1 gene. Mutations in this gene can lead to PRS superactivity, an X‑linked condition associated with excessive purine production.

Overactivity of PRPS1 increases PRPP availability, resulting in excess uric acid — a breakdown product of purines. Elevated uric acid can cause gout (arthritis due to uric acid crystals in joints) and kidney or bladder stones.

Increased PRPP availability may result from PRPP‑S superactivity or HPRT deficiency.

PRECICE® PRPP-S Assay Kit reaction principle

PRECICE® PRPP-S Assay Kit

Ref. #K0709-04-2

Kit Description

The PRECICE® PRPP-S Assay Kit is the first non‑radioactive solution for precise measurement of PRPP‑S activity in cellular lysates. This enzymatic assay uses highly active HGPRT and IMPDH enzymes to continuously monitor PRPP synthesis via NADH formation. Designed for research on purine salvage pathways and genetic disorders, the kit provides a fast, reliable, and user‑friendly protocol in a 96‑well plate format.

Contents of PRECICE® PRPP-S Assay Kit

Assay Principle

The assay continuously monitors PRPP synthesis through coupled enzymatic reactions:

  1. PRPP‑S catalyzes PRPP formation from ATP and ribose-5-phosphate.
  2. HGPRT converts PRPP and hypoxanthine (Hx) into IMP.
  3. IMPDH oxidizes IMP to XMP in the presence of NAD, producing NADH₂, measured spectrophotometrically at 340 nm.
#REF SIZE PRICE
#K0709-04-2 PRECICE® PRPP-S Assay Kit
10 mL of reaction mixture (half 96-well plate)		 515.00 €
515.00 €
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Updated on October 9th, 2025.

Kit is provided in stable lyophilized form and shipped without dry ice.

  User Manual (PDF)

Validated

Specific activity of PRPP‑S in human erythrocyte lysates measured with the kit is consistent with published data.

Validation graph of PRPP-S assay
RBC specimen 1 (n=4) 65 ± 2 nmol/h/mg Hb
RBC specimen 2 (n=4) 76 ± 12 nmol/h/mg Hb
RBC specimen 3 (n=4) 78 ± 10 nmol/h/mg Hb
Published data (Gordon et al., J. Inher. Metab. Dis. 10, 1987) 102 ± 20 nmol/h/mg Hb

Convenient

  • Non‑radioactive
  • Continuous monitoring
  • No sample preparation required — cell lysates are directly used

Fast

Analyze up to 6 samples in duplicate within 1 hour.

Principle Recap

PRPP‑S activity is quantified by monitoring NADH₂ formation at 340 nm after sequential HGPRT and IMPDH reactions.

Frequently Asked Questions

Find answers to common questions about the PRECICE® PRPP-S Assay Kit

The assay couples PRPP synthesis with HGPRT and IMPDH reactions. PRPP is converted to IMP, then oxidized to XMP, producing NADH₂ that is continuously monitored at 340 nm.

The kit quantifies PRPP in the range of 8–400 µM. Since intracellular PRPP is typically 0.1–1 µM, extracts should be concentrated 10–100 fold for accurate measurement.

PRPP is highly unstable. Use autoclaved solutions, precipitate cellular enzymes during extraction, and include positive controls to check for degradation.

A plate agitator, a plate reader with 340 nm filter or spectrophotometer, polypropylene tubes, and 0.2 µm PTFE syringe filters are required.

Up to 6 samples in duplicate can be analyzed simultaneously within 1 hour.

Have more questions? Our team is here to help.

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Scientific Works citing NOVOCIB PRPP-S Assay Kit:
  1. Direct stimulation of de novo nucleotide synthesis by O-GlcNAcylation 2024 L. Chen, Q. Zhou, P. Zhang, W. Tan, Y. Li, Z. Xu, J. Ma, G.M. Kupfer , Y. Pei, Q. Song 5, H. Pei Nature Chem Biol; 20(1):19-29
  2. Ablation of Atp5if1 impairs metabolic reprogramming and proliferation of T lymphocytes and compromises mouse survival I. Romero-Carraminana, S. Dominguez-Zorita, P.B. Esparza-Molto, J. M. Cuezva (2024) iScience 27, 109863
  3. Effect of hypoxia on purine metabolism in human skeletal muscle cells. (2021) Rivera-Pérez et al: Biotecnia / XXIII (2): 141-148
  4. PRPS-1 is a regulative for neuroprotection and cells regenerative proliferation (2018), Danielyan KE, Vardanyan R, Paronyan ZK , Barkhudaryants IM , Chailyan SG, Bisharyan MS. J Biomol Biochem Vol.2 No.2 6-10
  5. Comparison of human erythrocyte purine nucleotide metabolism and blood purine and pyrimidine degradation product concentrations before and after acute exercise in trained and sedentary subjects (2018) W. Dudzinska, M Suska, A Lubkowska, K Jakubowska, M Olszewska, K Safranow, D Chlubek
    J Physiol Sci 68(3):293-305.
  6. Cell cycle regulation of purine synthesis by phosphoribosyl pyrophosphate and inorganic phosphate (2013) Fridman; A. Saha; A. Chan; D.E. Casteel; R.B. Pilz; G.R. Boss Biochem J 454 (1): 91-99 A.