Recombinant Human Arylsulfatase B/ARSB Protein, CF

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

Recombinant Human Arylsulfatase B/ARSB Protein, CF Summary

• Details of Functionality: Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is >3,000 pmol/min/µg, as measured under the described conditions.
• Source: Chinese Hamster Ovary cell line, CHO-derived human Arylsulfatase B/ARSB protein
  Ser37-Met533, with a C-terminal 10-His tag
• Accession #: P15848
• N-terminal Sequence: Ser37
• Protein/Peptide Type: Recombinant Enzymes
• Gene: ARSB
• 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: 80 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, pH 6.5
  • Recombinant Human Arylsulfatase B/ARSB (rhARSB) (Catalog # 4415-SU)
  • Substrate: 4-Nitrocatechol Sulfate (4-PNCS) (Sigma, Catalog # N-7251)
  • NaOH (Sigma, Catalog # S-0899)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhARSB to 1 µg/mL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Combine 75 µL of 1 µg/mL rhARSB and 75 µL of 2 mM Substrate. Include a Substrate Blank containing 75 µL Assay Buffer and 75 µL Substrate.
  4. Incubate at 37 °C for 1 hour.
  5. Stop reaction by adding 150 µL of 0.2 M NaOH.
  6. Load 200 µL of reaction into a plate.
  7. Read at 510 nm (absorbance) in endpoint mode.
  8. 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:
  • rhARSB: 0.050 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 Human Arylsulfatase B/ARSB Protein, CF

• ARSB
• Arylsulfatase B
• ASB
• EC 3.1.6.12
• G4S
• MPS6
• N-acetylgalactosamine-4-sulfatase

Background

ARSB is a lysosomal enzyme of the sulfatase family that functions as an N-acetylgalactosamine-4-sulfatase (1). The enzyme is also active against chondroitin sulfate and dermatan sulfate (2). Deficiencies in ARSB activity are responsible for the lysosomal storage disease mucopolysaccharidosis Type VI, also known as Maroteaux-Lamy Syndrome (3).

1. Peters, C. et al. (1990) J. Biol. Chem. 265:3374.
2. O’Brien, J.S. et al. (1974) Biochem. Biophys. Res. Commun. 60:1170.
3. Wicker, G. et al. (1991) J. Biol. Chem. 266:21386.

Publications for Arylsulfatase B/ARSB (4415-SU)(1)

Publication using 4415-SU

• Bhattacharyya, S;O?Sullivan, I;Tobacman, J; N-Acetylgalactosamine-4-sulfatase (Arylsulfatase B) Regulates PD-L1 Expression in Melanoma by an HDAC3-Mediated Epigenetic Mechanism International Journal of Molecular Sciences 2024-05-28 [PMID: 38892038] (Bioassay, In vivo assay, Human, Mouse)

Bioinformatics

• Gene Symbol: ARSB
• Uniprot:
  • P15848()