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Recombinant Human Sulfamidase/SGSH Protein, CF

Catalog #: 8380-SU Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
8380-SU-020
Recombinant Human Sulfamidase/SGSH Protein Enzyme Activity Diagram.
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Sulfamidase/SGSH Protein, CF Summary

Product Specifications

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.
Activity
Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is >0.3 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 #
P51688
N-terminal Sequence
Analysis
Arg23
Predicted Molecular Mass
55 kDa
SDS-PAGE
55-63 kDa, under reducing conditions.

Product Datasheets

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8380-SU

Product Datasheet
COA
SDS

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.

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.

Assay Procedure

Materials
  • Assay Buffer: 50 mM MES, pH 5.5
  • Recombinant Human Sulfamidase/SGSH (rhSGSH) (Catalog # 8380-SU)
  • Substrate: 4-Nitrocatechol Sulfate (PNCS) (Sigma, Catalog # N7251), 100 mM stock in deionized water
  • Sodium Hydroxide (NaOH) (Sigma, Catalog # 221465), 2 M stock in deionized water
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  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 in microtubes. Include a Substrate Blank with 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 to each tube.
  6. Load 200µL from each stopped reaction and Substrate Blank vial into a plate.
  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) (Sigma, Catalog # N15553).

Per Well:
  • rhSGSH: 2 µg
  • Substrate: 5 mM

      • Scientific Data

      Enzyme Activity View Larger

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

      SDS-PAGE View Larger

      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.

      Reconstitution Calculator

      Reconstitution Calculator

      The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 (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.
      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

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