Recombinant Human NAGLU Protein, CF

R&D Systems | Catalog # 7096-GH

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human NAGLU Protein (7096-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

CHO

Accession Number

P54802

Applications

Enzyme Activity
View all alpha-N-acetylglucosaminidase/NAGLU Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human alpha-N-acetylglucosaminidase/NAGLU protein
Met1-Trp743, 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

Asp24

Predicted Molecular Mass

81 kDa

SDS-PAGE

70-95 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze 4-Nitrophenyl-N-acetyl-alpha -D-glucosaminide.
The specific activity is >900 pmol/min/μg, as measured under the decribed conditions.

Scientific Data Images for Recombinant Human NAGLU Protein, CF

Recombinant Human NAGLU Protein Enzyme Activity Diagram.

Recombinant Human NAGLU Protein, CF (Catalog # 7096-GH) degrades heparan sulfate by hydrolysis of terminal GlcNAc resides in N-acetyl-alpha-D-glucosaminides of heparan sulfate.

Recombinant Human NAGLU Protein Enzyme Activity.

Recombinant Human NAGLU Protein, CF (Catalog # 7096-GH) is measured by its ability to hydrolyze 4-Nitrophenyl-N-acetyl-alpha -D-glucosaminide.

Recombinant Human NAGLU Protein SDS-PAGE.

2 μg/lane of Recombinant Human NAGLU Protein (Catalog # 7096-GH) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 70-95 kDa.

Formulation, Preparation, and Storage

7096-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: alpha-N-acetylglucosaminidase/NAGLU

Human lysosomal alpha -N-acetylglucosaminidase is a hydrolase that catalyses the removal of terminal alpha ‑N‑acetylglucosamine residues from heparan sulfate and heparin (1). Defects in this gene are the cause of mucopolysaccharidosis type IIIB (MPS‑IIIB), also known as Sanfilippo syndrome B (2, 3, 4). Mucopolysaccharidosis types IIIA, C, and D are caused by mutations in other genes involved in the lysosomal degradation of heparan sulfate. Continuous lysosomal accumulation of heparan sulfate results in the clinical onset of disease, which is typified by severe central nervous system degeneration (5). Mucopolysaccharidosis type III differs from other mucopolysaccharidoses in that patients usually exhibit mild somatic changes with minimal skeletal abnormalities (6).

References

  1. Yogalingam, G. et al. (2000) Biochim. Biophy. Acta 1502:415.
  2. Zhao, H.G. et al. (1996) Proc. Natl. Acad. Sci. U.S.A. 93:6101.
  3. Schmidtchen, A. et al. (1998) Am. J. Hum. Genet. 62:64.
  4. Weber, B. et al. (1999) Eur. J. Hum. Genet. 7:34.
  5. Beesley, C.E. et al. (2005) J. Inherit. Metab. Dis. 28:759.
  6. Yogalingam, G. and Hopwood, J.J. (2001) Hum. Mutat. 18:264.

Alternate Names

alphaNacetylglucosaminidase, NAG, NAGLU, UFHSD1

Entrez Gene IDs

4669 (Human); 27419 (Mouse)

Gene Symbol

NAGLU

UniProt

P54802

Additional alpha-N-acetylglucosaminidase/NAGLU Products

Product Documents for Recombinant Human NAGLU 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 NAGLU Protein, CF

For research use only

Citations for Recombinant Human NAGLU Protein, CF

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Protocols

View specific protocols for Recombinant Human NAGLU Protein, CF (7096-GH):

Materials
  • Assay Buffer: 0.1 M Sodium Citrate, 0.25 M NaCl, pH 4.5
  • Recombinant Human alpha ‑N‑acetylglucosaminidase/NAGLU (rhNAGLU) (Catalog # 7096-GH)
  • Substrate: 4-Nitrophenyl-N-acetyl-alpha -D-glucosaminide (Sigma, Catalog # N-8759), 5 mM stock in deionized water
  • NaOH, 0.2 M stock in deionized water
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhNAGLU to 3 ng/μL in Assay Buffer.
  2. Dilute Substrate to 3 mM in Assay Buffer.
  3. In a plate combine 50 μL of rhNAGLU and 50 μL of 3 mM Substrate. Include a Substrate Blank containing 50 μL Assay Buffer and 50 μL of 3 mM Substrate.
  4. Seal plate and incubate at 37 °C for 20 minutes.
  5. Add 100 μL of 0.2 M NaOH to each well to stop the reaction and develop the color.
  6. Read absorbance in endpoint mode at 402 nm.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Abs* (OD) x well volume (L) x 1012 pmol/mol
Inc. time (min) x epsilon ** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Using the extinction coefficient 17700 M-1cm-1
     ***Using the path correction 0.6 cm

Per Well:
  • rhNAGLU: 0.150 μg
  • Substrate: 750 μM

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