Recombinant Mouse Iduronate 2-sulfatase Protein, CF
Recombinant Mouse Iduronate 2-sulfatase Protein, CF Summary
Product Specifications
Thr36-Pro552 with an N-terminal signal sequence and a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
|
Assay Procedure
- 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
- Dilute rmIDS to 20 µg/mL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- 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.
- Incubate at 37 °C for 24 hours.
- Stop reaction by adding 300 µL of 0.2 M NaOH to reaction tubes.
- Load 200 µL from each reaction tube in duplicate into a 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).
- rmIDS: 1.0 µg
- Substrate: 1 mM
Reconstitution Calculator
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 # 2449-SU).
- Parenti, G. et al. (1997) Curr. Opin. Genet. & Dev. 7:386.
- 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.
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