Recombinant Human Sulfamidase/SGSH Protein, CF

R&D Systems | Catalog # 8380-SU

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human Sulfamidase/SGSH Protein (8380-SU)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

Sf 21 (baculovirus)

Accession Number

P51688

Applications

Enzyme Activity
View all Sulfamidase/SGSH Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Sulfamidase/SGSH protein
Arg23-Leu502 with a C-terminal 6-His tag

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Arg23

Predicted Molecular Mass

55 kDa

SDS-PAGE

55-63 kDa, under reducing conditions.

Activity

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.

Scientific Data Images for Recombinant Human Sulfamidase/SGSH Protein, CF

Recombinant Human Sulfamidase/SGSH Protein Enzyme Activity Diagram.

Recombinant Human Sulfamidase/SGSH Protein, CF (Catalog # 8380-SU) hydrolyzes N-linked sulfate group from the non-reducing terminal glucosaminide residues of heparan sulfate.

Recombinant Human Sulfamidase/SGSH Protein SDS-PAGE.

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 at 55-63 kDa.

Formulation, Preparation, and Storage

8380-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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

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 (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.

References

  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.

Alternate Names

HSS, MPS3A, SFMD, SGSH

Entrez Gene IDs

6448 (Human); 27029 (Mouse)

Gene Symbol

SGSH

UniProt

P51688

Additional Sulfamidase/SGSH Products

Product Documents for Recombinant Human Sulfamidase/SGSH Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human Sulfamidase/SGSH Protein, CF

For research use only

Citations for Recombinant Human Sulfamidase/SGSH Protein, CF

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Protocols

View specific protocols for Recombinant Human Sulfamidase/SGSH Protein, CF (8380-SU):

Materials
  • 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

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