Recombinant Mouse Sulfamidase/SGSH Protein, CF

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Product Details

Summary
Reactivity MuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Mouse Sulfamidase/SGSH Protein, CF Summary

Details of Functionality
Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is >0.4 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
Arg23
Protein/Peptide Type
Recombinant Enzymes
Gene
Sgsh
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
56 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
60 kDa, reducing conditions
Publications
Read Publications using
2969-SU in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
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
  1. Dilute rmSGSH to 40 µg/mL in Assay Buffer.
  2. Dilute Substrate to 20 mM in Assay Buffer.
  3. 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.
  4. Incubate at 37 °C for 72 hours.
  5. Stop reaction by adding 300 µL of 0.2 M NaOH to the reaction vials.
  6. Load in a plate 200 µL in duplicate from each reaction vial.
  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:
  • rmSGSH: 2 µg
  • Substrate: 5 mM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse Sulfamidase/SGSH Protein, CF

  • heparan sulfate sulfatase
  • HSS
  • HSSN-sulphoglucosamine sulphohydrolase
  • MPS3A
  • MPS3Asulphamidase
  • N-sulfoglucosamine sulfohydrolase
  • SFMD
  • SFMDEC 3.10.1.1
  • SGSH
  • Sulfamidase
  • Sulfoglucosamine sulfamidase
  • Sulphamidase

Background

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.

  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.

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Publications for Sulfamidase/SGSH (2969-SU)(2)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 1 application: Bioassay.


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Bioassay
(2)
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(1)
Mouse
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Bioinformatics

Gene Symbol Sgsh
Entrez
Uniprot