Recombinant Human Iduronate 2-Sulfatase Protein, CF Summary
Ser26-Pro550, with a C-terminal 10-His tag
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|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.
- 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
- Dilute rhIDS to 20 ng/µL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- Combine equal volumes of 20 ng/µL rhIDS and 2 mM Substrate. Include a Substrate Blank containing Assay Buffer and Substrate.
- Incubate at 37 °C for 24 hours.
- Stop reaction by adding equivalent volume (total volume of step 3) of 0.2 M NaOH to reaction tubes.
- Load 200 µL from each reaction tube into the plate.
- Read at 510 nm (absorbance) in endpoint mode.
- 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
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.
- Wilson, P.J. et al. (1990) Proc. Natl. Acad. Sci. USA 87:8531.
- Parenti, G. et al. (1997) Curr. Opin. Genet. & Dev. 7:386.
- 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.
Citations for Recombinant Human Iduronate 2-Sulfatase Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
Citations: Showing 1 - 2
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Secondary storage of dermatan sulfate in Sanfilippo disease.
Authors: Lamanna WC, Lawrence R, Sarrazin S, Esko JD
J. Biol. Chem., 2011;286(9):6955-62.
Sample Types: Cell Lysates
Pharmacoproteomics of a metalloproteinase hydroxamate inhibitor in breast cancer cells: dynamics of membrane type 1 matrix metalloproteinase-mediated membrane protein shedding.
Authors: Butler GS, Dean RA, Tam EM, Overall CM
Mol. Cell. Biol., 2008;28(15):4896-914.
Sample Types: Whole Cells
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