Recombinant Human Iduronate 2-Sulfatase Protein, CF

R&D Systems | Catalog # 2449-SU

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Iduronate 2-Sulfatase Protein (2449-SU)
  • 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

NS0

Accession Number

P22304

Applications

Enzyme Activity
View all Iduronate 2-Sulfatase/IDS Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Iduronate 2-Sulfatase/IDS protein
Ser26-Pro550, with a C-terminal 10-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

Ser26

Predicted Molecular Mass

61 kDa

SDS-PAGE

Multiple bands between 74-91 kDa, reducing conditions

Activity

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.

Formulation, Preparation, and Storage

2449-SU
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: Iduronate 2-Sulfatase/IDS

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.

References

  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.

Alternate Names

SIDS

Entrez Gene IDs

3423 (Human); 15931 (Mouse)

Gene Symbol

IDS

UniProt

P22304

Additional Iduronate 2-Sulfatase/IDS Products

Product Documents for Recombinant Human Iduronate 2-Sulfatase Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Iduronate 2-Sulfatase Protein, CF

For research use only

Citations for Recombinant Human Iduronate 2-Sulfatase Protein, CF

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Protocols

View specific protocols for Recombinant Human Iduronate 2-Sulfatase Protein, CF (2449-SU):

Materials
  • 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

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