Recombinant Human QSOX1/Quiescin Q6 Protein, CF

R&D Systems | Catalog # 9209-QS

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human QSOX1/Quiescin Q6 Protein (9209-QS)
  • 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

E. coli

Accession Number

O00391

Applications

Enzyme Activity
View all QSOX1/Quiescin Q6 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human QSOX1/Quiescin Q6 protein
Ser33-Ala546, with an N-terminal Met and 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

Met

Predicted Molecular Mass

58 kDa

SDS-PAGE

55 kDa, reducing conditions

Activity

Measured by its ability to produce hydrogen peroxide during the oxidation of Dithiothreitol (DTT).
The specific activity is >1,200 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

9209-QS
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: QSOX1/Quiescin Q6

Sulfhydryl Oxidase-1 (QSOX1) is an approximately 80 kDa enzyme that contains thioredoxin and sulfhydryl oxidase domains (1-3). It is synthesized with a C-terminal transmembrane segment, but soluble secreted forms can be generated by alternative splicing or proteolytic shedding within the Golgi (4). Within the region encompassing both enzymatic domains and the central region, human QSOX1 shares 79% aa sequence identity with mouse and rat QSOX1. It plays a role nascent protein folding by mediating disulfide oxidation (4-6). This activity is required for Laminin incorporation into the extracellular matrix (7). QSOX1 is up-regulated in many cancers and supports tumor cell proliferation and invasion (1, 8).

References

  1. Lake, D.F. and D.O. Faigel (2014) Antioxid. Redox. Signal. 21:485.
  2. Kodali, V.K. and C. Thorpe (2010) Antioxid. Redox Signal. 13:1217.
  3. Coppock, D.L. et al. (1998) Genomics 54:460.
  4. Rudolf, J. et al. (2013) Biochem. J. 454:181.
  5. Heckler, E.J. et al. (2008) Biochemistry 47:4955.
  6. Alon, A. et al. (2012) Nature 488:414.
  7. Ilani, T. et al. (2013) Science 341:74.
  8. Katchman, B.A. et al. (2011) Mol. Cancer Res. 9:1621.

Long Name

Quiescin Q6 Sulfhydryl Oxidase 1

Alternate Names

HQSOX, QSCN6, Quiescin Q6, Sulfhydryl Oxidase-1

Entrez Gene IDs

5768 (Human); 104009 (Mouse); 84491 (Rat)

Gene Symbol

QSOX1

UniProt

O00391

Additional QSOX1/Quiescin Q6 Products

Product Documents for Recombinant Human QSOX1/Quiescin Q6 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.

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Product Specific Notices for Recombinant Human QSOX1/Quiescin Q6 Protein, CF

For research use only

Related Research Areas

Oxidases and Oxygenases
Redox Enzymes

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Protocols

View specific protocols for Recombinant Human QSOX1/Quiescin Q6 Protein, CF (9209-QS):

Materials
  •  Assay Buffer: 50 mM Sodium Phosphate, pH 7.5
  • Recombinant Human QSOX1/Quiescin Q6 (rhQSOX1) (Catalog # 9209-QS)
  • Coupling Enzyme: Horseradish Peroxidase (HRP) (250-330 U/mg) (Sigma, Catalog # P8375), 250 units/mL stock in 0.1 M Sodium Phosphate, pH 8.0
  • Substrate Component 1: Dithiothreitol (DTT) (Amresco, Catalog # 0281), 1 M stock in deionized water
  • Substrate Component 2:  Amplex® Ultra Red (AUR) (Invitrogen, Catalog # A36006), 10 mM stock in DMSO
  • F15 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhQSOX1 to 1 ng/μL in Assay Buffer.
  2. Prepare a Substrate Mixture containing 100 µM AUR, 2 units/mL HRP, and 300 µM DTT in Assay Buffer. Make sure to add DTT to the Substrate Mixture right before loading the plate.
  3. Load 50 μL of 1 ng/μL rhQSOX1 into the plate, and start the reaction by adding 50 μL of Substrate Mixture. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of Substrate Mixture.
  4. Read at excitation and emission wavelengths of 544 nm and 590 nm in kinetic mode for 5 minutes. Note: A cutoff must be set at a wavelength of 570 nm.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using a fluorescent standard prepared by incubating 50 µM AUR, 1 unit/mL HRP and 150 µM DTT and a curve of Hydrogen Peroxide (Sigma, Catalog # H1009) in Assay Buffer. Use this oxidized AUR curve to determine the conversion factor.

Per Well:
  • rhQSOX1: 0.05 μg
  • DTT: 150 µM
  • HRP: 1 unit/mL
  • AUR: 50 µM

FAQs

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