E. coli-derived human D-Amino Acid Oxidase protein Arg2-Leu347 with N-terminal Met and 6-His tag
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Predicted Molecular Mass
39 kDa, under reducing conditions
Measured by its ability to produce hydrogen peroxide during the oxidative deamination of D-amino acids. The specific activity is >1000 pmol/min/μg, as measured under the described conditions.
Recombinant Human D-Amino Acid Oxidase His-tag Protein, CF Scientific Data Examples
Recombinant Human D-Amino Acid Oxidase His-tag Protein SDS-PAGE
1 μg/lane of Recombinant Human D-Amino Acid Oxidase His-tag (Catalog # 10116-DO) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing a band at 39 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 dry ice or equivalent. 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.
Background: D-Amino Acid Oxidase
D-amino acid oxidase (DAO) is an FAD-dependent enzyme that catalyzes the strict stereoselective oxidative deamination of D-amino acids. Human DAO is an 80 kDa stable homodimer of monomers in head to head conformation (1). Each monomer has two domains: a substrate-binding domain containing the active site and responsible for oligomerization and the FAD-binding domain (2). Mammalian DAO is primarily expressed in kidney, liver and brain, but also in small intestine and neutrophilic leukocytes (3). Human DAO differs in biophysical characteristics compared to other species due to its weakly bound cofactor, modulation by the regulatory protein pLG72 (4), and relatively low activity on its physiological substrates: D-serine, D-Cys, and D-DOPA (5). D-serine is a key neuromodulator and co-agonist of N-methyl-D-aspartate receptors (NMDAR), receptors that are critically involved in learning and memory. Abnormal levels of D-serine are associated with chronic neurodegeneration and misregulation of DAO. While many variants of DAO are implicated in neurological disease, a specific DAO gene mutation resulting in an enzyme with impaired activity causes dominantly inherited familial amyotrophic lateral sclerosis (ALS) (6). Only variants in the DAO gene are significantly associated with the clinical outcome of ALS patients (7). In Alzheimer's disease (AD), DAO expression is higher and correlates with severity of cognitive impairment (8) while inhibition resulted in improved cognitive function in AD patients (9). Likewise, as with AD, increased expression of DAO with concomitant decreased D-serine levels is found in schizophrenia with beneficial results observed upon therapeutic delivery of D-serine (10). DAO inhibition has also been shown to modulate chronic pain (11, 12). DAO is a pharmaceutical target for the treatment of neurological therapeutic disease and pain (13, 14).
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Kawazoe, T. et al. (2007) Biochem. Biophys. Res. Commun. 355:385.
Mitchell, J. et al. (2010) Proc. Natl. Acad. Sci. U.S.A. 107:7556.
Cirulli, E.T. et al. (2015) Science 347:1436.
Lin, C. H. et al. (2017) J. Alzhemier's Dis. 56:959.
Lin, C.H. et al. (2014) Curr. Pharm. Des. 20:5169.
Heresco-Levy, U. et al. (2005) Biol. Psychiatry 57:577.
Zhou, W.J. et al. (2010) J. Pharmacol. Exp. Ther. 332:248.
Gong, N. et al. (2011) J. Pharmacol. Exp. Ther. 336:282.
Szilagyi, B. et al. (2018) Expert Opin. Drug Discov. 13:973.
Khangura, R.K. et al. (2019) Korean J. Physiol. Pharmacol. 23:1.
Which Brands are Currently Available on bio‑techne.com? R&D Systems, Tocris Bioscience and ProteinSimple branded products are available to purchase through bio‑techne.com. ProteinSimple branded instruments are available to quote. ACD branded products will be available on bio‑techne.com in the near future. Novus Biologicals branded products are not currently available on bio‑techne.com and can be found at novusbio.com.
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