Recombinant Human Glucosamine (N-acetyl)-6-Sulfatase, CF

R&D Systems | Catalog # 2484-SUC

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human Glucosamine (N-acetyl)-6-Sulfatase (2484-SUC)
  • 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

CHO

Accession Number

P15586

Applications

Enzyme Activity
View all Glucosamine (N-acetyl)-6-Sulfatase/GNS Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Glucosamine (N-acetyl)-6-Sulfatase/GNS protein
Val37-Leu552 & Thr44-Leu552, both with a C-terminal 10-His tag

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.

N-terminal Sequence Analysis

Val37 & Thr44

Predicted Molecular Mass

59 & 60 kDa

SDS-PAGE

94 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS).
The specific activity is >250 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 5 using 2484-SUC in the following applications:

Formulation, Preparation, and Storage

2484-SUC
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
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: Glucosamine (N-acetyl)-6-Sulfatase/GNS

A member of the sulfatase family, GNS is required for the lysosomal degradation of the glycosaminoglycans (GAG) heparan sulfate and keratan sulfate (1, 2). It hydrolyzes the 6-sulfate group of the N-acetyl-D-glucosamine 6-sulfate units of the GAG. GNS deficiency results in mucopolysaccharidosis type IIID (MPS IIID or Sanfilippo D Syndrome), an inborn error leading to lysosomal accumulation of heparan sulfate. MPS IIID has profound mental deterioration, hyperactivity, and relatively mild somatic manifestations. The deduced amino acid sequence of human GNS consists of a signal peptide (residues 1-36) and a mature chain (residues 37-552) that may be further processed into N-terminal and C-terminal fragments (3). rhGNS corresponds to the single chain and has sulfatase activity described in Activity Assay Protocol.

References

  1. Parenti, G. et al. (1997) Curr. Opin. Genet. & Dev. 7:386.
  2. Neufeld, E.F. and J. Muenzer (2001) in The Metabolic and Molecular Basis of Inherited Disease, Scriver, C.R. et al. (eds.) pp. 3421, New York, McGraw-Hill.
  3. Robertson, D.A. et al. (1992) Biochem. J. 288:539.

Alternate Names

Glucosamine6Sulfatase, GNS

Entrez Gene IDs

2799 (Human)

Gene Symbol

GNS

UniProt

P15586

Additional Glucosamine (N-acetyl)-6-Sulfatase/GNS Products

Product Documents for Recombinant Human Glucosamine (N-acetyl)-6-Sulfatase, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Glucosamine (N-acetyl)-6-Sulfatase, CF

For research use only

Citations for Recombinant Human Glucosamine (N-acetyl)-6-Sulfatase, CF

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    Verified Customer | Posted 01/23/2017
    Recombinant Human Glucosamine (N-acetyl)-6-Sulfatase, CF 2484-SUC

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Protocols

View specific protocols for Recombinant Human Glucosamine (N-acetyl)-6-Sulfatase, CF (2484-SUC):

Materials
  • Assay Buffer: 50 mM Sodium Acetate, 250 mM NaCl, pH 5.0
  • Recombinant Human Glucosamine (N-acetyl)‑6-Sulfatase/GNS (rhGNS) (Catalog # 2484-SUC)
  • Substrate: 4-Nitrocatechol Sulfate (PNCS) (Sigma, Catalog # N-7251), 100 mM stock in deionized water
  • 0.2 M NaOH
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhGNS to 2 µg/mL in Assay Buffer.
  2. Dilute PNCS substrate to 4 mM in Assay Buffer.
  3. Combine 75 µL of 2 µg/mL rhGNS and 75 µL 4 mM PNCS. Include a Substrate Blank containing Assay Buffer in place of rhGNS.
  4. Incubate at 37 °C for 2 hours.
  5. Stop reactions and develop color by adding 150 µL of 0.2 M NaOH to each reaction.
  6. Load into the wells of a clear 96-well plate 200 µL from each reaction.
  7. Read at 510 nm (absorbance) in endpoint mode.
  8. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Abs* (OD) x Conversion Factor** (pmole/OD)
Incubation time (min) x amount of enzyme (µg)

     *Adjusted for Substrate Blank

     **Derived using calibration standard p-Nitrocatechol (Sigma, Catalog # N15553).

Per Well:
  • rhGNS: 0.1 µg
  • Substrate: 1 mM

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