Recombinant Mouse Sulfamidase/SGSH Protein, CF

Catalog # Availability Size / Price Qty
2969-SU-020

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Recombinant Mouse Sulfamidase/SGSH Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is >0.25 pmol/min/µg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse Sulfamidase/SGSH protein
Arg23-Leu502, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Arg23
Predicted Molecular Mass
56 kDa
SDS-PAGE
60 kDa, reducing conditions

Product Datasheets

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2969-SU

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 50 mM MES, pH 5.5
  • Recombinant Mouse Sulfamidase/SGSH (mSGSH) (Catalog # 2969-SU)
  • Substrate: 4-Nitrocatechol Sulfate (PNCS), 100 mM stock in deionized water
  • Sodium Hydroxide (NaOH), 2 M stock in deionized water
  • Clear 96-well Plate
  • Plate Reader
  1. Dilute rmSGSH to 40 µg/mL in Assay Buffer.
  2. Dilute Substrate to 20 mM in Assay Buffer.
  3. Combine 150 µL of 40 µg/mL rmSGSH and 150 µL of 20 mM Substrate. Include a Substrate Blank containing 150 µL of Assay Buffer and 150 µL of 20 mM Substrate.
  4. Incubate at 37 °C for 72 hours.
  5. Stop reactions and Substrate Blank by adding 300 µL of 0.2 M NaOH.
  6. Load in a plate 200 µL of each reaction and Substrate Blank.
  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).

Per Well:
  • rmSGSH: 2 µg
  • Substrate: 5 mM
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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.

References
  1. Diez-Roux, G. and Ballabio, A. (2005) Annu. Rev. Genomics Hum. Genet. 6:355.
  2. Gliddon, B.L. et al. (2004) Mol. Genet. Metab. 83:239.
Entrez Gene IDs
6448 (Human); 27029 (Mouse)
Alternate Names
heparan sulfate sulfatase; HSS; HSSN-sulphoglucosamine sulphohydrolase; MPS3A; MPS3Asulphamidase; N-sulfoglucosamine sulfohydrolase; SFMD; SFMDEC 3.10.1.1; SGSH; Sulfamidase; Sulfoglucosamine sulfamidase; Sulphamidase

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.

2 Citations: Showing 1 - 2
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  1. 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, 2018-02-27;8(1):3681.
    Species: Mouse
    Sample Types:
    Applications: Bioassay
  2. Secondary storage of dermatan sulfate in Sanfilippo disease.
    Authors: Lamanna WC, Lawrence R, Sarrazin S, Esko JD
    J. Biol. Chem., 2010-12-30;286(9):6955-62.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay

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