Recombinant Human Iduronate 2-Sulfatase Protein, CF

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

Recombinant Human Iduronate 2-Sulfatase Protein, CF Summary

• Details of Functionality: Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is > 1.0 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Iduronate 2-Sulfatase/IDS protein
  Ser26-Pro550, with a C-terminal 10-His tag
• Accession #: P22304
• N-terminal Sequence: Ser26
• Protein/Peptide Type: Recombinant Enzymes
• Gene: IDS
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 61 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: Multiple bands between 74-91 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 visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Assay Procedure:
  • Assay Buffer: 50 mM Sodium Acetate, 100 mM NaCl, pH 5.0
  • Recombinant Human Iduronate 2‑Sulfatase/IDS (rhIDS) (Catalog # 2449-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 rhIDS to 20 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Combine equal volumes of 20 ng/µL rhIDS and 2 mM Substrate. Include a Substrate Blank containing Assay Buffer and Substrate.
  4. Incubate at 37 °C for 24 hours.
  5. Stop reaction by adding equivalent volume (total volume of step 3) of 0.2 M NaOH to reaction tubes.
  6. Load 200 µL from each reaction tube into the 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:
  • rhIDS: 1.0 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 Iduronate 2-Sulfatase Protein, CF

• Alpha-L-iduronate sulfate sulfatase
• EC 3.1.6.13
• IDS
• iduronate 2-sulfatase 14 kDa chain
• iduronate 2-sulfatase 42 kDa chain
• iduronate 2-sulfatase
• idursulfase
• MPS2
• S
• SIDS

Background

As a member of the sulfatase family, IDS is required for the lysosomal degradation of the glycosaminoglycans (GAG) heparan sulfate and dermatan sulfate (2, 3). It hydrolyzes the 2-sulfate group of the L-iduronate 2-sulfate units of the GAG. The IDS deficiency results in mucopolysaccharidosis II (MPS II or Hunter syndrome), an X-linked inborn error leading to lysosomal accumulation of the GAG and its excretion in urine. MPS II has a wide spectrum of clinical manifestations ranging from mild to severe. The deduced amino acid sequence of human IDS consists of a signal peptide (residues 1‑25), a pro peptide (residues 26‑33) and a mature chain (residues 34‑550) that may be further processed into the 42 kDa chain (residues 34‑455) and the 14 kDa chain (residues 456‑550) (1). rhIDS corresponds to the single chain and has sulfatase activity described above.

1. Wilson, P.J. et al. (1990) Proc. Natl. Acad. Sci. USA 87:8531.
2. Parenti, G. et al. (1997) Curr. Opin. Genet. & Dev. 7:386.
3. Neufeld, E.F. and Muenzer, J. (2001) in The Metabolic and Molecular Basis of Inherited Disease, Scriver, C.R. et al. (eds.) pp. 3421, New York, McGraw-Hill.

Publications for Iduronate 2-Sulfatase/IDS (2449-SU)(2)

Publications using 2449-SU

• Lamanna WC, Lawrence R, Sarrazin S, Esko JD Secondary storage of dermatan sulfate in Sanfilippo disease. J. Biol. Chem., 2010-12-30;286(9):6955-62. 2010-12-30 [PMID: 21193389] (Bioassay, Human)
• Butler GS, Dean RA, Tam EM, Overall CM Pharmacoproteomics of a metalloproteinase hydroxamate inhibitor in breast cancer cells: dynamics of membrane type 1 matrix metalloproteinase-mediated membrane protein shedding. Mol. Cell. Biol., 2008-05-27;28(15):4896-914. 2008-05-27 [PMID: 18505826] (Bioassay, Human)

Bioinformatics

• Gene Symbol: IDS
• Uniprot:
  • P22304()