Recombinant Human beta-Glucuronidase/GUSB Protein, CF

R&D Systems | Catalog # 6144-GH

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human beta-Glucuronidase/GUSB Protein (6144-GH)
  • 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

NS0

Accession Number

AAA52561

Applications

Enzyme Activity
View all beta-Glucuronidase/GUSB Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human beta-Glucuronidase/GUSB protein
Met1-Thr651, with a C-terminal 6-His tag

Purity

>95%, 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

Leu23

Predicted Molecular Mass

73 kDa

SDS-PAGE

75-85 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze 4-methylumbelliferyl-beta -D-glucuronide.
The specific activity is >4,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6144-GH
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: beta-Glucuronidase/GUSB

Human beta -Glucuronidase (EC 3.2.1.31) encoded by the GUSB gene is a lysosomal hydrolase involved in the stepwise degradation of glucuronic acid-containing glycosaminoglycans that include heparan sulfate, chondroitin sulfate and hyaluronan (1). The enzyme is only active on the glucuronic acid of the non-reducing end. The native protein has been reported as a tetrameric glycoprotein composed of identical subunits (1, 2). Mutations in the GUSB gene are linked to mucopolysaccharidosis type VII (3). Accumulation of partially degraded glycosaminoglycans, with glucuronic acid residues at the non-reducing termini, are usually found in the lysosomes of patients with the disease (3). It has also been reported that this enzyme may contribute to the depletion of chondroitin from cartilage and thereby facilitate the damage of joints in rheumatoid arthritis (4).

References

  1. Shipley, J.M. et al. (1993) Am. J. Hum. Genet. 52:517.
  2. Oshima, A, et al. (1987) Proc. Natl. Acad. Sci. USA 84:685.
  3. Bell, C.E. Jr. et al. (1977) J. Clin. Invest. 59:97.
  4. Ortutay, Z. et al. (2003) Arthritis Rheum. 48:2163.

Alternate Names

Beta-G1, MPS7

Entrez Gene IDs

2990 (Human)

Gene Symbol

GUSB

UniProt

AAA52561

Additional beta-Glucuronidase/GUSB Products

Product Documents for Recombinant Human beta-Glucuronidase/GUSB Protein, 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 beta-Glucuronidase/GUSB Protein, CF

For research use only

Citations for Recombinant Human beta-Glucuronidase/GUSB Protein, CF

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Protocols

View specific protocols for Recombinant Human beta-Glucuronidase/GUSB Protein, CF (6144-GH):

Materials
  • Assay Buffer: 100 mM Sodium Acetate, pH 3.5
  • Recombinant Human beta ‑Glucuronidase/GUSB (rhGUSB) (Catalog # 6144-GH)
  • Substrate: 4-Methylumbelliferyl beta -D-glucuronide (Sigma, Catalog # M9130), 50 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhGUSB to 4 ng/μL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load into a plate 50 μL of 4 ng/μL rhGUSB and start the reaction by adding 50 μL of 2 mM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 2 mM Substrate.
  4. Read plate at excitation and emission wavelengths of 365 nm and 445 nm, respectively, in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 4-Methylumbelliferone (4-MU) (Sigma, Catalog # M1381).

Per Well:
  • rhGUSB: 0.200 μg
  • Substrate: 1 mM

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