Recombinant Human Tissue alpha-L-Fucosidase/FUCA1, CF

R&D Systems | Catalog # 7039-GH

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Tissue alpha-L-Fucosidase/FUCA1 (7039-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

P04066

Applications

Enzyme Activity
View all Tissue alpha-L-Fucosidase/FUCA1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Tissue alpha-L-Fucosidase/FUCA1 protein
Gln32-Lys466 with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Gln32

Predicted Molecular Mass

51 kDa

SDS-PAGE

55-60 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic substrate 4-methylumbelliferyl-alpha -L-fucopyranoside.
The specific activity is >4,800 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

7039-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: Tissue alpha-L-Fucosidase/FUCA1

Fucosylated glycoconjugates play numerous roles in biological processes, such as development, apoptosis and the immune responses, and are involved in the pathology of inflammation, cancer and cystic fibrosis (1, 2, 3, 4). Tissue alpha ‑L‑Fucosidase, also known as FUCA1, is a lysosomal enzyme responsible for hydrolyzing alpha ‑L‑fucose moieties from glycolipids and oligosaccharides (5). The mature enzyme in the liver has a molecular weight of 200 kDa and exists in the native state as a tetramer (6). Defects in FUCA1 are the cause of fucosidosis (7, 8, 9), an autosomal recessive lysosomal storage disease characterized by the accumulation of fucose-containing glycolipids and glycoproteins in various tissues. Serum alpha ‑L‑fucosidase has been identified as a useful marker for oral cancer (10).

References

  1. Hiraishi, K. et al. (1993) Glycobiology 3:381.
  2. Solter, D. and Knowles, B.B. (1978) Proc. Natl. Acad. Sci. USA 75:5565.
  3. Becker, D.J. and Lowe, J.B. (2003) Glycobiology 13:41R.
  4. Liu, T.W. et al. (2009) Proc. Natl. Acad. Sci. USA 106:14581.
  5. Fukushima, H. et al. (1985) Proc. Natl. Acad. Sci. USA 82:1262.
  6. Alhadeff, J.A. et al. (1975) J. Biol. Chem. 250:7106.
  7. O’Brien, J.S. et al. (1987) Enzyme 38:45.
  8. Durand, P. et al. (1969) J. Pediatr. 75:665.
  9. Johnson, K. and Dawson, G. (1985) Biochem. Biophys. Res. Commun. 133:90.
  10. Shah, M. et al. (2008) Cancer 113:336.

Alternate Names

FUCA1, Tissue alphaLFucosidase

Entrez Gene IDs

2517 (Human); 71665 (Mouse); 24375 (Rat)

Gene Symbol

FUCA1

UniProt

P04066

Additional Tissue alpha-L-Fucosidase/FUCA1 Products

Product Documents for Recombinant Human Tissue alpha-L-Fucosidase/FUCA1, 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 Tissue alpha-L-Fucosidase/FUCA1, CF

For research use only

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Protocols

View specific protocols for Recombinant Human Tissue alpha-L-Fucosidase/FUCA1, CF (7039-GH):

Materials
  • Assay Buffer: 50 mM Sodium Acetate, 5 mM MgCl2, pH 4.5
  • Recombinant Human Tissue alpha ‑L‑Fucosidase/FUCA1 (rhFUCA1) (Catalog # 7039-GH)
  • Substrate: 4-Methylumbelliferyl-alpha -L-fucopyranoside (Research Products International Corp, Catalog # M65200), 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 rhFUCA1 to 2 ng/μL in Assay Buffer.
  2. Dilute Substrate to 1.6 mM in Assay Buffer.
  3. Load into a plate 50 μL of 2 ng/μL rhFUCA1, and start the reaction by adding 50 μL of 1.6 mM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 1.6 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:
  • rhFUCA1: 0.100 μg
  • Substrate: 0.8 mM

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