Recombinant Mouse Iduronate 2-sulfatase Protein, CF

R&D Systems | Catalog # 2486-SU

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Iduronate 2-sulfatase Protein (2486-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

NP_034628

Applications

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

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Iduronate 2-Sulfatase/IDS protein
Thr36-Pro552 with an N-terminal signal sequence and 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

Thr36

Predicted Molecular Mass

60 kDa

SDS-PAGE

77 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

2486-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, Iduronate 2-Sulfatase encoded by the IDS gene is required for the lysosomal degradation of the glycosaminoglycans (GAG) heparan sulfate and dermatan sulfate (1, 2). 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 their excretion in urine. MPS II has a wide spectrum of clinical manifestations ranging from mild to severe. The mouse IDS has sulfatase activity described in the Activity Assay Protocol. In addition, recombinant human IDS is also available (Catalog # http://www.rndsystems.com/product_results.aspx?k=2449-SU">2449-SU).

References

  1. Parenti, G. et al. (1997) Curr. Opin. Genet. & Dev. 7:386.
  2. Neufeld, E.F. and J. Muenzer (2001) in The Metabolic and Molecular Basis of Inherited Disease, Scriver, C.R. et al. (eds.) p. 3421, New York, McGraw-Hill.

Alternate Names

SIDS

Entrez Gene IDs

3423 (Human); 15931 (Mouse)

Gene Symbol

IDS

UniProt

NP_034628

Additional Iduronate 2-Sulfatase/IDS Products

Product Documents for Recombinant Mouse 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 Mouse Iduronate 2-sulfatase Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Mouse Iduronate 2-sulfatase Protein, CF (2486-SU):

Materials
  • Assay Buffer: 50 mM NaOAc, pH 5.0
  • Recombinant Mouse Iduronate 2‑Sulfatase/IDS (rmIDS) (Catalog # 2486-SU)
  • Substrate: 4-Nitrocatechol Sulfate (4-PNCS) (Sigma, Catalog # N-7251)
  • Sodium Hydroxide (NaOH) (Sigma, Catalog # S-0899)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: Spectra-Max Plus by Molecular Devices) or equivalent
  1. Dilute rmIDS to 20 µg/mL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Combine 150 µL of 20 µg/mL rmIDS and 150 µL of 2 mM Substrate. Include a Substrate Blank containing Assay Buffer and Substrate only.
  4. Incubate at 37 °C for 24 hours.
  5. Stop reaction by adding 300 µL of 0.2 M NaOH to reaction tubes.
  6. Load 200 µL from each reaction tube in duplicate 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:
  • rmIDS: 1.0 µg
  • Substrate: 1 mM

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