Recombinant Human Dopamine beta-Hydroxylase Protein, CF

R&D Systems | Catalog # 7376-AO

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human Dopamine beta-Hydroxylase Protein (7376-AO)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

CHO

Accession Number

P09172

Applications

Enzyme Activity
View all Dopamine beta-Hydroxylase Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Dopamine beta-Hydroxylase protein
Met1-Gly617, with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Leu37 & Ser40

Predicted Molecular Mass

66 kDa

SDS-PAGE

65-85 kDa, reducing conditions

Activity

Measured by its ability to convert tyramine to octopamine.
The specific activity is >7,000 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 4 using 7376-AO in the following applications:

Formulation, Preparation, and Storage

7376-AO
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
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.

Background: Dopamine beta-Hydroxylase

Dopamine beta -Hydroxylase (DBH), also known as dopamine beta -monooxygenase, belongs to the copper type II, ascorbate-dependent monooxygenase family. DBH is found within the neurosecretory vesicles of adrenal medullae and the large dense‑cored synaptic vesicles of the sympathetic nervous system as both membrane‑associated and soluble forms (1, 2). It catalyzes the conversion of dopamine to noradrenaline in sympathetic neurons, making it an important enzyme for catecholamine biosynthesis (3, 4). Mutations in the DBH gene that result in low DBH activity are a cause of noradrenaline deficiency (5). This recombinant human DBH is the secreted, soluble form.

References

  1. Dhawan, S. et al. (1987) J. Biol. Chem. 262:1869.
  2. Saxena, A. and P.J. Fleming (1983) J. Biol. Chem. 258:4147.
  3. Levin, E.Y. et al. (1960) J. Biol. Chem. 235:2080.
  4. Friedman, S. and S. Kaufman (1965) J. Biol. Chem. 240:4763.
  5. Kim, C.H. et al. (2002) Am. J. Med. Genet. 108:140.

Alternate Names

DBH, DBM, Dopamine betaHydroxylase, DOPBHY

Entrez Gene IDs

1621 (Human); 13166 (Mouse); 25699 (Rat)

Gene Symbol

DBH

UniProt

P09172

Additional Dopamine beta-Hydroxylase Products

Product Documents for Recombinant Human Dopamine beta-Hydroxylase Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Dopamine beta-Hydroxylase Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human Dopamine beta-Hydroxylase Protein, CF (7376-AO):

Materials
  • Assay Buffer: 0.1 M NaOAc, 0.5 µM CuCl2, pH 5.0
  • Recombinant Human Dopamine beta ‑Hydroxylase (rhDBH) (Catalog # 7376-AO)
  • Substrate Component 1: Tyramine (Sigma, Catalog # T2879), 100 mM stock in deionized water
  • Substrate Component 2: N,N-Dimethyl-p-phenylenediamine (DMPD) (Sigma, Catalog # D4139), 500 mM stock in deionized water
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhDBH to 1 ng/µL in Assay Buffer.
  2. Dilute both substrate components to 40 mM respectively in Assay Buffer.
  3. Combine equivalent volumes of substrate components for substrate mixture.
  4. Load 50 µL of the diluted rhDBH into a clear plate, and start the reaction by adding 50 µL of the Substrate mixture to wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate Mixture without any rhDBH.
  5. Read in kinetic mode for 5 minutes at an absorbance of 515 nm.
  6. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) 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 5200 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Well:
  • rhDBH: 0.050 μg
  • Tyramine: 10 mM
  • DMPD: 10 mM

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