Recombinant T. maritima alpha-L-Fucosidase Protein, CF

R&D Systems | Catalog # 6556-GH

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Key Product Details

  • R&D Systems E. coli-derived Recombinant T. maritima alpha-L-Fucosidase Protein (6556-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

E. coli

Accession Number

NP_228118

Applications

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

Product Specifications

Source

E. coli-derived t. maritima alpha-L-Fucosidase protein
Ile2-Glu449, with an N-terminal Met and 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

Met

Predicted Molecular Mass

53 kDa

SDS-PAGE

43-50 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic substrate 4-methylumbelliferyl-alpha -L-fucopyranoside.
The specific activity is >1,600 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 5 using 6556-GH in the following applications:

Formulation, Preparation, and Storage

6556-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-L-Fucosidase

Fucosylated glycoconjugates play numerous roles in biological events, including development and apoptosis (1, 2), and are involved in the pathology of inflammation, cancer, and cystic fibrosis (3, 4). Fucosidases are generally used for studying fucosylated glycans. With a Kcat/Km value of 160,000 M-1s-1, the alpha -L-fucosidases of Thermotoga maritima efficiently hydrolyzes 4-nitrophenyl fucoside (5). The enzyme is the closest bacterial relative of mammalian fucosidase with 38% identity to its human homologue. It is thought to remove alpha ‑1,2‑ and alpha -1,4-linked fucosyl side chains from algal fucoidan (5), while the activity on alpha -1,3- and alpha ‑1,6‑ linked fucose has not been tested. The enzyme assembles as a hexamer and displays a two-domain fold, with Asp224 and Glu266 being critical for enzyme activity (6).

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. Shah, M. et al. (2008) cancer 113:338.
  4. Becker, D.J. and Lowe, J.B. (2003) Glycobiology 13:41R.
  5. Tarling, C.A. et al. (2003) J. Biol. Chem. 278:47394.
  6. Sulzenbacher, G. et al. (2004) J. Biol. Chem. 279:13119.

Alternate Names

alphaLFucosidase

Entrez Gene IDs

897238 (T. maritima)

Gene Symbol

TM_RS01570

UniProt

NP_228118

Additional alpha-L-Fucosidase Products

Product Documents for Recombinant T. maritima alpha-L-Fucosidase 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 T. maritima alpha-L-Fucosidase Protein, CF

For research use only

Customer Reviews for Recombinant T. maritima alpha-L-Fucosidase Protein, CF (1)

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    Verified Customer | Posted 10/06/2020
    Recombinant T. maritima alpha-L-Fucosidase Protein, CF 6556-GH

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Protocols

View specific protocols for Recombinant T. maritima alpha-L-Fucosidase Protein, CF (6556-GH):

Materials
  • Assay Buffer: 50 mM MES, 200 mM NaCl, pH 5.5
  • Recombinant T. maritima alpha ‑L‑Fucosidase (rT. maritima alpha -L-Fucosidase) (Catalog # 6556-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 rT. maritima alpha -L-Fucosidase to 2 ng/μL in Assay Buffer.
  2. Dilute Substrate to 400 μM in Assay Buffer.
  3. Load into a plate 50 μL of 2 ng/μL rT. maritima alpha -L-Fucosidase, and start the reaction by adding 50 μL of 400 μM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 400 μM 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 (Sigma, Catalog # M1381).

Per Well:
  • rT. maritima alpha -L-Fucosidase: 0.100 μg
  • Substrate: 200 μM

FAQs

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