Recombinant Mouse Sulfamidase/SGSH Protein, CF
Recombinant Mouse Sulfamidase/SGSH Protein, CF Summary
Product Specifications
Arg23-Leu502, with a C-terminal 10-His tag
Analysis
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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.
2969-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 MES, pH 5.5
- Recombinant Mouse Sulfamidase/SGSH (mSGSH) (Catalog # 2969-SU)
- Substrate: 4-Nitrocatechol Sulfate (PNCS) (Sigma, Catalog # N-7251), 100 mM stock in deionized water
- Sodium Hydroxide (NaOH) (Sigma, Catalog # 221465), 2 M stock in deionized water
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rmSGSH to 40 µg/mL in Assay Buffer.
- Dilute Substrate to 20 mM in Assay Buffer.
- Mix 150 µL of 40 µg/mL of rmSGSH and 150 µL of 20 mM of Substrate. Include a Substrate Blank with 150 µL of Assay Buffer and 150 µL of Substrate.
- Incubate at 37 °C for 72 hours.
- Stop reaction by adding 300 µL of 0.2 M NaOH to the reaction vials.
- Load in a plate 200 µL in duplicate from each reaction vial.
- 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:- rmSGSH: 2 µg
- Substrate: 5 mM
Reconstitution Calculator
Background: Sulfamidase/SGSH
Also known as N-sulfoglucosamine sulfohydrolase and heparan N-sulfatase, Sulfamidase/SGSH is an important member of the sulfatase family involved in the degradation of heparan sulfate (1). The SGSH deficiency results in mucopolysaccharidosis type IIIA (MPS IIIA, Sanfilippo A syndrome), an autosomal recessive lysosomal storage disease characterized by neurological dysfunction but relatively mild somatic manifestations (2). The deduced amino acid sequence of mouse SGSH consists of a signal peptide (residues 1-22) and a mature chain (residues 23-502) (1). Recombinant Mouse Sulfamidase/SGSH corresponds to the mature chain and has sulfatase activity as described in the Activity section.
- Diez-Roux, G. and Ballabio, A. (2005) Annu. Rev. Genomics Hum. Genet. 6:355.
- Gliddon, B.L. et al. (2004) Mol. Genet. Metab. 83:239.
Citations for Recombinant Mouse Sulfamidase/SGSH 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.
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Citations: Showing 1 - 2
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Fast, sensitive method for trisaccharide biomarker detection in mucopolysaccharidosis type 1
Authors: E Makino, H Klodnitsky, J Leonard, J Lillie, TC Lund, J Marshall, J Nietupski, PJ Orchard, WP Miller, C Phaneuf, D Tietz, ML Varban, M Donovan, A Belenki
Sci Rep, 2018;8(1):3681.
Species: Mouse
Sample Types:
Applications: Bioassay -
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.
Species: Human
Sample Types: Cell Lysates
Applications: Bioassay
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