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Recombinant Human UPP1 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human UPP1 Protein, CF Summary

Details of Functionality
Measured by its ability to catalyze the phosphorolysis of uridine to uracil and ribose-1-phosphate. The specific activity is >9,000 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived human Uridine Phosphorylase 1/UPP1 protein
Ala2-Ala310, with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Met
Protein/Peptide Type
Recombinant Enzymes
Gene
UPP1
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
35 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
32-34 kDa, reducing conditions

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Tris, NaCl, DTT and Glycerol.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Assay Procedure
  • Assay Buffer: 0.1 M Potassium Phosphate, pH 7.4
  • Recombinant Human Uridine Phosphorylase 1/UPP1 (rhUPP1) (Catalog # 7234-UP)
  • Substrate: Uridine (Sigma, Catalog # U3003), 100 mM stock in deionized water
  • UV plate (Costar, Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhUPP1 to 2 ng/μL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load into a plate 50 μL of 2 ng/μL rhUPP1, and start the reaction by adding 50 μL of 2 mM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 2 mM Substrate.
  4. Read plate at a wavelength of 280 nm (bottom read) in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x -1 x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

     *Adjusted for Substrate Blank 
     **Using the extinction coefficient 1370 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Well:
  • rhUPP1: 0.1 μg
  • Substrate: 1 mM

Notes

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.


This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human UPP1 Protein, CF

  • EC 2.4.2.3
  • UDPRASE
  • UDRPASE
  • UP
  • UPASE 1
  • UPASE
  • UPP
  • UPP1
  • UPUPase 1
  • UrdPase 1
  • uridine phosphorylase 1
  • uridine phosphorylase

Background

Uridine Phophorylase 1 (UPP1) belongs to the family of pentosyltransferases. UPP1 is expressed in the cytosol of most tissues and at elevated levels in many tumors (1, 2). It plays an important role in the pyrimidine salvage pathway through its catalysis of the reversible phosphorolysis of uridine to uracil (1, 2). UPP1 is important for the homeostatic regulation of intracellular and plasma uridine concentrations (3). UPP1 also plays an important role in the activation of 5-fluorouracil, an anti-tumor drug (4). The high level of UPP1 expression in some tumors makes it a potential prognostic factor for some cancers, such as oral squamous cell carcinoma (5).
  1. Krenitsky, T. A. et al. (1964) J. Biol. Chem. 239:805.
  2. Cass, C. E. and Paterson, A. R. P. (1972) J. Biol. Chem. 247:3314.
  3. Connolly, G.P. and Duley, J. A. (1999) Trends Pharmacol. Sci. 20:218.
  4. Cao, D. and Pizzorno, G. (2004) Drugs Today (Barc) 40:431.
  5. Miyashita, H. et al. (2002) Cancer 94:2959.

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Bioinformatics

Gene Symbol UPP1
Uniprot