Recombinant Human Kynurenine 3-Monooxygenase/KMO Protein, CF

R&D Systems | Catalog # 8050-KM

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human Kynurenine 3-Monooxygenase/KMO Protein (8050-KM)
  • 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

Sf 21 (baculovirus)

Accession Number

O15229

Applications

Enzyme Activity
View all Kynurenine 3-Monooxygenase/KMO Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Kynurenine 3-Monooxygenase/KMO protein
Asp2-Leu441, with an N-terminal Met and 6-His tag

Purity

>80%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

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

N-terminal Sequence Analysis

Inconclusive results, intact N-terminus verified by anti-poly-His Western

Predicted Molecular Mass

51 kDa

SDS-PAGE

40-45 kDa, reducing conditions

Activity

Measured by the consumption of NADPH by hydroxylation of L-kynurenine to 3-hydroxy-kynurenine.
The specific activity is >290 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

8050-KM
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Brij-35.
Shipping 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: Kynurenine 3-Monooxygenase/KMO

Kynurenine 3-monooxygenase (KMO), also known as kynurenine 3-hydroxylase, is a part of the kynurenine pathway of tryptophan degradation (1). KMO catalyzes the NADPH- and flavin adenine dinucleotide (FAD)-dependent 3-hydroxylation of kynurenine to 3-hydroxykynurenine (3-HK). 3-HK is neurotoxic via the generation of hydrogen peroxide (2) and through the excitotoxic effects of its downstream metabolite quinolinic acid (3). The levels of 3-HK and quinolinic acid are increased in the brain with Alzheimer's disease and Huntington's disease (1). Inhibition of KMO was shown to reverse cognitive and motor deficits in mouse models of those diseases via an increase in neuroprotective kynurenic acid (4). KMO is found in the mitochondrial outer membrane of microglial cells in the brain and dendritic cells and macrophages in the periphery (1). This recombinant human KMO was expressed as a C-terminally truncated protein.

References

  1. Schwarcz, R. et al. (2012) Nat. Rev. Neurosci. 13:465.
  2. Okuda, S. et al. (1996) Proc. Natl. Acad. Sci. 93:12553.
  3. Stone, T.W. and M.N. Perkins. (1981) Eur. J. Pharmacol. 72:411.
  4. Zwilling, D. et al. (2011) Cell 145:863.

Alternate Names

KMO, Kynurenine 3Monooxygenase

Entrez Gene IDs

8564 (Human); 98256 (Mouse); 59113 (Rat)

Gene Symbol

KMO

UniProt

O15229

Additional Kynurenine 3-Monooxygenase/KMO Products

Product Documents for Recombinant Human Kynurenine 3-Monooxygenase/KMO Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human Kynurenine 3-Monooxygenase/KMO Protein, CF

For research use only

Citations for Recombinant Human Kynurenine 3-Monooxygenase/KMO Protein, CF

Customer Reviews for Recombinant Human Kynurenine 3-Monooxygenase/KMO Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human Kynurenine 3-Monooxygenase/KMO Protein, CF and earn rewards!

Have you used Recombinant Human Kynurenine 3-Monooxygenase/KMO Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human Kynurenine 3-Monooxygenase/KMO Protein, CF (8050-KM):

Materials
  • Assay Buffer: 50 mM Sodium Phosphate, 0.1% (w/v) Brij-35, pH 7.5
  • Recombinant Human Kynurenine 3-Monooxygenase/KMO (rhKMO) (Catalog # 8050-KM)
  • beta -Nicotinamide adenine dinucleotide phosphate reduced, tetrasodium salt ( beta -NADPH) (Sigma, Catalog # N7505), 10 mM stock in deionized water
  • L-Kynurenine (Tocris, (Catalog # 4393), 40 mM stock in 80 mM HEPES, pH 8.0
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhKMO to 20 ng/μL in Assay Buffer.
  2. Prepare a Reaction Mixture containing 400 μM beta -NADPH and 600 μM L-Kynurenine in Assay Buffer.
  3. In a plate, load 50 μL of 20 ng/μL rhKMO, and start the reaction by adding 50 μL of Reaction Mixture.
  4. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of Reaction Mixture.
  5. Read at an absorbance of 340 nm in kinetic mode for 10 minutes.
  6. 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 6270 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Well:

  • rhKMO: 1 μg
  • beta -NADPH: 200 μM
  • L-Kynurenine: 300 μM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes