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Recombinant Human NAGLU Protein, CF

Catalog #: 7096-GH
2 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
7096-GH-020
Recombinant Human NAGLU Protein Enzyme Activity Diagram.
3 Images
Product Details
Citations (2)
FAQs
Reviews
Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human NAGLU Protein, CF Summary

Product Specifications

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.
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.
Source
Chinese Hamster Ovary cell line, CHO-derived human alpha-N-acetylglucosaminidase/NAGLU protein
Met1-Trp743, with a C-terminal 6-His tag
Accession #
P54802
N-terminal Sequence
Analysis
Asp24
Predicted Molecular Mass
81 kDa
SDS-PAGE
70-95 kDa, reducing conditions

Product Datasheets

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7096-GH

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.

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.

Assay Procedure

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

Scientific Data

Enzyme Activity View Larger

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.

Enzyme Activity View Larger

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

SDS-PAGE View Larger

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.

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: 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.
Entrez Gene IDs
4669 (Human); 27419 (Mouse)
Alternate Names
alphaNacetylglucosaminidase; alpha-N-acetylglucosaminidase; EC 3.2.1.50; MPS3B; MPS-IIIB; N-acetyl-alpha-glucosaminidase; N-acetylglucosaminidase, alpha; NAG; NAGalpha-N-acetylglucosaminidase; NAGLU; UFHSD; UFHSD1

Citations for Recombinant Human NAGLU Protein, 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.

2 Citations: Showing 1 - 2
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  1. Structure and Mechanism of Staphylococcus aureus TarS, the Wall Teichoic Acid ?-glycosyltransferase Involved in Methicillin Resistance
    PLoS Pathog, 2016-12-14;12(12):e1006067.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Structure and mechanism of Staphylococcus aureus TarM, the wall teichoic acid alpha-glycosyltransferase.
    Authors: Sobhanifar S, Worrall L, Gruninger R, Wasney G, Blaukopf M, Baumann L, Lameignere E, Solomonson M, Brown E, Withers S, Strynadka N
    Proc Natl Acad Sci U S A, 2015-01-26;112(6):E576-85.
    Species: Bacteria - Staphylococcus aureus
    Sample Types: Whole Cells
    Applications: Bioassay

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