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

Catalog # Availability Size / Price Qty
2484-SUC-010
R&D Systems Recombinant Proteins and Enzymes
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Citations (1)
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Reviews (1)

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

Product Specifications

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.
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.
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
Accession #
N-terminal Sequence
Analysis
Val37 & Thr44
Predicted Molecular Mass
59 & 60 kDa
SDS-PAGE
94 kDa, reducing conditions

Product Datasheets

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2484-SUC

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.

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.

Assay Procedure

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
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: 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.
Entrez Gene IDs
2799 (Human)
Alternate Names
EC 3.1.6; EC 3.1.6.14; G6Sglucosamine-6-sulfatase; glucosamine (N-acetyl)-6-sulfatase; Glucosamine6Sulfatase; Glucosamine-6-Sulfatase; GNS; MGC21274; N-acetylglucosamine-6-sulfatase

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

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Oligosaccharides mapping of nitrous acid degraded heparin through UHPLC-HILIC/WAX-MS
    Authors: T Zhang, S Xie, Z Wang, R Zhang, Q Sun, X Liu, L Chi, JP Li, H Li, T Tan
    Carbohydr Polym, 2020;231(0):115695.
    Species: Human
    Sample Types: Whole Cells
    Applications: Cell Culture

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Recombinant Human Glucosamine (N-acetyl)-6-Sulfatase, CF
By Derek Moen on 01/23/2017
Application: Enzymatic activity in vitro
Reason for Rating: Provides very reproducible activity measurements along with long-term stability - serves as an ideal positive control for both activity assays and SDS-PAGE applications. Did not demonstrate in-vivo intracellular uptake.