Recombinant Human Sulfamidase/SGSH Protein, CF

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Recombinant Human Sulfamidase/SGSH Protein, CF (Catalog # 8380-SU) hydrolyzes N-linked sulfate group from the non-reducing terminal glucosaminide residues of heparan sulfate.
2 μg/lane of Recombinant Human Sulfamidase/SGSH Protein (Catalog # 8380-SU) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human Sulfamidase/SGSH Protein, CF Summary

Details of Functionality
Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is >0.125 pmol/min/μg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Sulfamidase/SGSH protein
Arg23-Leu502 with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Arg23
Protein/Peptide Type
Recombinant Enzymes
Gene
SGSH
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
55 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
55-63 kDa, under reducing conditions.
Publications
Read Publication using
8380-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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • Assay Buffer: 50 mM MES, pH 5.5
  • Recombinant Human Sulfamidase/SGSH (rhSGSH) (Catalog # 8380-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 (Catalog # DY990)
  • Plate Reader
  1. Dilute rhSGSH to 40 µg/mL in Assay Buffer.
  2. Dilute PNCS to 20 mM in Assay Buffer.
  3. Combine 150 µL of 40 µg/mL of rhSGSH and 150 µL of 20 mM of PNCS. Include a Substrate Blank containing 150 µL of Assay Buffer and 150 µL of 20 mM of PNCS.
  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 into a plate 200 µL from each stopped reaction and Substrate Blank.
  7. Read plate 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 4-Nitrocatechol (PNC).

Per Well:
  • rhSGSH: 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 Human 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 (HS) (1). Different from the HS specific endosulfatases that remove sulfate from internal GlcNAc residues (2), SGSH removes sulfate group from the non-reducing end glucosamine residues on HS. 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 (3). Human SGSH shows 88.6% sequence identity with that of mouse sequence.
      1. DiezRoux, G. and Ballabio, A. et al. (2005) Annu. Rev. Genomics Hum. Genet. 6:355.
      2. Morimoto-Tomita, M. et al. (2002) J. Biol. Chem. 277:49175.
      3. Blanch, L. et al. (1997) Hum. Mol. Genet. 6:787.

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      Publications for Sulfamidase/SGSH (8380-SU)(1)

      We have publications tested in 1 confirmed species: N/A.

      We have publications tested in 1 application: Bioassay.


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      Bioinformatics

      Gene Symbol SGSH
      Uniprot