Recombinant Mouse Arylsulfatase G/ARSG Protein, CF

• Reactivity: Mu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Mouse Arylsulfatase G/ARSG Protein, CF Summary

• Details of Functionality: Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is > 1,500 pmol/min/µg, as measured under the described conditions.
• Source: Chinese Hamster Ovary cell line, CHO-derived mouse Arylsulfatase G/ARSG protein
  Gly17-Val525, with a C-terminal 10-His tag
  Accession # Q3TYD4
• Accession #: Q3TYD4
• N-terminal Sequence: Gly17
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Arsg
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 57 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 65 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris and NaCl.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Assay Procedure:
  • Assay Buffer: 50 mM MES, 250 mM NaCl, pH 5.5
  • Recombinant Mouse Arylsulfatase G/ARSG (rmARSG) (Catalog # 4600-SU)
  • Substrate: 4-Nitrocatechol Sulfate (PNCS) (Sigma, Catalog # N-7251), 100 mM in deionized water
  • Sodium Hydroxide, 1 M stock in deionized water
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rmARSG to 1 µg/mL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Mix 75 μL of 1 µg/mL of rmARSG and 75 µL of 2 mM of Substrate in duplicate. Create a Substrate Blank with 75 µL Assay Buffer and 75 µL Substrate in duplicate. Incubate at 37 °C for 1 hour.
  4. Stop reaction by adding 150 µL of 0.2 M sodium hydroxide to the reaction vials.
  5. Load into a clear 96-well plate 200 µL from each reaction vial.
  6. Read at 510 nm (absorbance) in endpoint mode.
  7. Calculate specific activity:

  Specific Activity (pmol/min/µg) = (Adjusted Abs* (OD) x Conversion Factor** (pmol/OD)) / (Incubation time (min) x amount of enzyme (µg))

  *Adjusted for Substrate Blank
  **Derived using calibration standard P-Nitrocatechol (PNC) (Sigma, Catalog # N15553).

  Per Well:
  • rmARSG: 0.05 µg
  • Substrate: 0.5 mM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse Arylsulfatase G/ARSG Protein, CF

• ARSG
• Arylsulfatase G
• ASG
• EC 3.1.6
• EC 3.1.6.-
• EC 3.1.6.4
• KIAA1001ASG

Background

ARSG is a member of the aryl sulfatase family of enzymes with a high degree of amino acid sequence similarity to ARSA and ARSB. ARSG mRNA is expressed in a ubiquitous manner, being detected in most adult and embryonic tissues of the mouse (1).

1. Ferrante, F. et al. (2002) Eur. J. Hum. Genetics 10:813.

Bioinformatics

• Gene Symbol: Arsg
• Uniprot:
  • Q3TYD4()