Recombinant Human NAGLU Protein, CF
Recombinant Human NAGLU Protein, CF Summary
Met1-Trp743, with a C-terminal 6-His tag
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.
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.
|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.
- 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
- Dilute rhNAGLU to 3 ng/μL in Assay Buffer.
- Dilute Substrate to 3 mM in Assay Buffer.
- 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.
- Seal plate and incubate at 37 °C for 20 minutes.
- Add 100 μL of 0.2 M NaOH to each well to stop the reaction and develop the color.
- Read absorbance in endpoint mode at 402 nm.
- 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
- rhNAGLU: 0.150 μg
- Substrate: 750 μM
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).
- Yogalingam, G. et al. (2000) Biochim. Biophy. Acta 1502:415.
- Zhao, H.G. et al. (1996) Proc. Natl. Acad. Sci. U.S.A. 93:6101.
- Schmidtchen, A. et al. (1998) Am. J. Hum. Genet. 62:64.
- Weber, B. et al. (1999) Eur. J. Hum. Genet. 7:34.
- Beesley, C.E. et al. (2005) J. Inherit. Metab. Dis. 28:759.
- Yogalingam, G. and Hopwood, J.J. (2001) Hum. Mutat. 18:264.
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.
Citations: Showing 1 - 2
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Structure and Mechanism of Staphylococcus aureus TarS, the Wall Teichoic Acid ?-glycosyltransferase Involved in Methicillin Resistance
PLoS Pathog, 2016;12(12):e1006067.
Sample Types: Whole Cells
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;112(6):E576-85.
Species: Bacteria - Staphylococcus aureus
Sample Types: Whole Cells
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