E. coli-derived human DHODH protein Thr31 - Arg395 with N-terminal Met and 6-His tag
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Predicted Molecular Mass
41 kDa, reducing conditions
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.
Recombinant Human Dihydroorotate Dehydrogenase Protein, CF Scientific Data Examples
Recombinant Human Dihydroorotate Dehydrogenase Protein Binding Activity
Recombinant Human DHODH (Catalog # 10062-DD) is measured by its reduction of 2,6-dichloroindophenol during oxidation of dihydroorotate.
Recombinant Human Dihydroorotate Dehydrogenase Protein SDS-PAGE
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.
Formulation, Preparation and Storage
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
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
Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
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 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
Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
Dilute rhDHODH to 0.4 µg/mL in Assay Buffer.
Prepare Substrate Mixture containing 2 mM L-Dihydroorotic acid, 0.2 mM Decylubiquinone and 0.12 mM DPIP in Assay Buffer.
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.
Immediately read plate in kinetic mode for 5 minutes at 600 nm (absorbance).
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.
rhDHODH: 0.02 µg
L-Dihydroorotic acid: 1 mM
Decylubiquinone: 0.1 mM
DPIP: 0.06 mM
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).
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Lewis, T. A. et al. (2016) ACS Med. Chem. Lett. 7:1112.
Koundinya, M. et al. (2018) Cell Chem. Biol. 21:705.
Herrmann, M. L. et al. (2000) Immunopharmacology. 47:273.
Li, E. K. et al. (2004) Clin. Ther. 26:447.
Warnke, C. et al. (2013) Clin. Neurol. Neurosurg. 115:S90.
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Wang, Q. Y. et al. (2011) J. Virol. 85:6548.
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Fang, J. et al. (2012) Biosci. Rep. 32:631.
Entrez Gene IDs
1723 (Human); 56749 (Mouse); 65156 (Rat)
DHODH, DHOdehase, Dihydroorotate Oxidase, EC 126.96.36.199, EC 188.8.131.52, POADS, URA1, dihydroorotate dehydrogenase, dihydroorotate dehydrogenase, mitochondrial, human complement of yeast URA1
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