Recombinant Human Dihydroorotate Dehydrogenase Protein, CF

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Recombinant Human Dihydroorotate Dehydrogenase Protein Binding Activity
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Product Details

Recombinant Human Dihydroorotate Dehydrogenase Protein, CF Summary

Product Specifications

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Measured by its reduction of 2,6-dichloroindophenol during the oxidation of dihydroorotate. The specific activity is >15000 pmol/min/μg, as measured under the described conditions.
E. coli-derived human DHODH protein
Thr31 - Arg395
with N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Predicted Molecular Mass
41 kDa
41 kDa, reducing conditions

Product Datasheets

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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

  • Assay Buffer: 50 mM Tris, 150 mM KCl, 0.1% Triton® X-100, pH 8.0
  • Recombinant Human DHODH His-tag (rhDHODH) (Catalog # 10062-DD)
  • L-Dihydroorotic acid (Sigma, Catalog # D7128), 40 mM stock in DMF
  • Decylubiquinone (Sigma, Catalog # D7911), 20 mM stock in DMSO
  • 2,6-Dichloroindophenol sodium salt hydrate (DPIP) (Sigma, Catalog # D1878), 2 mM in Absolute Ethanol
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhDHODH to 0.4 µg/mL in Assay Buffer.
  2. Prepare Substrate Mixture containing 2 mM L-Dihydroorotic acid, 0.2 mM Decylubiquinone and 0.12 mM DPIP in Assay Buffer.
  3. Load 50 µL of 0.4 µg/mL rhDHODH to plate, and start the reaction by adding 50 µL of Substrate Mixture.  Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of Substrate Mixture.
  4. Immediately read plate in kinetic mode for 5 minutes at 600 nm (absorbance).
  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 21,000 M-1 cm-1.
***Using the path correction 0.32 cm.
Note: the output of many spectrophotometers is in mOD.

Per Well:
  • rhDHODH: 0.02 µg
  • L-Dihydroorotic acid: 1 mM
  • Decylubiquinone: 0.1 mM
  • DPIP: 0.06 mM

Scientific Data

Binding Activity Recombinant Human Dihydroorotate Dehydrogenase Protein Binding Activity View Larger

Recombinant Human DHODH (Catalog # 10062-DD) is measured by its reduction of 2,6-dichloroindophenol during oxidation of dihydroorotate.

SDS-PAGE Recombinant Human Dihydroorotate Dehydrogenase Protein SDS-PAGE View Larger

2 μg/lane of Recombinant Human DHODH was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing a band at 41 kDa.

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Background: DHODH

Dihydroorotate dehydrogenase (DHODH) is a rate-limiting enzyme in the pyrimidine de novo pathway that converts dihydroorotate to orotate. DHODHs have a monomeric structure composed of a large C-terminal a/b barrel and a small N-terminal helical domain (1). There are two main classes of DHODHs based on similarity, preference of substrate, and subcellular location (1,2). Within Class 2, structural differences can be exploited for selective targeting (3,4).  Human DHODH belongs to Class 2 and is a monomeric, mitochondrial, FMN-dependent enzyme (2). It is ubiquitously expressed in most tissue. As DHODH catalyzes de novo pyrimidine synthesis for synthesis of DNA/RNA essential to rapidly proliferating cells, DHODH is currently a target for treatment of cancer (5-8). It has also been successfully targeted in treatment for rheumatoid arthritis (9-10), multiple sclerosis (11-12), viral infection (13-14), microbial infectious diseases such as malaria (4, 15) and gastrointestinal disease (3, 16), and antifungal infection (17). Mutations in the DHODH resulting in functional defects cause Miller syndrome, also known as postaxial acrofacial dystosis syndrome (POADS) (18).

  1. Liu, S. et al. (2000) Structure. 8:25.
  2. Reis, R. A. G. et al. (2017) Arch. Biochem. Biophys. 632:175.
  3. Copeland, R. A. et al. (2000) J. Biol. Chem. 275:33373.
  4. Singh, A. et al. (2017) Eur. J. Med. Chem. 125:640.
  5. Baumann, P. et al. (2009) Mol. Cancer Ther. 8:366.
  6. Zhu, S. et al. (2013) PLoS One. 8:e71555.
  7. Lewis, T. A. et al. (2016) ACS Med. Chem. Lett. 7:1112.
  8. Koundinya, M. et al. (2018) Cell Chem. Biol. 21:705.
  9. Herrmann, M. L. et al. (2000) Immunopharmacology. 47:273.
  10. Li, E. K. et al. (2004) Clin. Ther. 26:447.
  11. Warnke, C. et al. (2013) Clin. Neurol. Neurosurg. 115:S90.
  12. O'Connor, P. et al. (2016) Neurology 86:920.
  13. Wang, Q. Y. et al. (2011) J. Virol. 85:6548.
  14. Smee, D. F. et al. (2012) Antivir. Chem. Chemother. 22:263.
  15. Skerlj, R. T. et al. (2011) ACS Med. Chem. Lett. 2:708.
  16. Ohishi, T. et al. (2018) Helicobacter 23:e12470.
  17. Oliver, J. D. et al. (2016) Proc. Natl. Acad. Sci. USA 113:12809.
  18. Fang, J. et al. (2012) Biosci. Rep. 32:631.
Long Name
Dihydroorotate Dehydrogenase
Entrez Gene IDs
1723 (Human); 56749 (Mouse); 65156 (Rat)
Alternate Names
DHOdehase; DHODH; dihydroorotate dehydrogenase; dihydroorotate dehydrogenase, mitochondrial; Dihydroorotate Oxidase; EC; EC; human complement of yeast URA1; POADS; URA1


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