Because N-, O-glycans and glycolipids are frequently fucosylated at terminal sites, fucose is often found to be essential for sugar epitope and lectin ligand generation. Well known fucose containing structures include Lewis structures and ABO blood group antigens. Lewis structures are key elements involved in leukocyte homing and extravasation process and thus are essential for lymphocyte maturation and natural defense functions. Fucose containing glycans also play essential roles in cell signaling and development. So far, more than 10 Fucosyltransferases have been cloned from the human genome (1). FUT1 and FUT2 are alpha 1‑2 Fucosyltransferases and are responsible for ABO blood group antigen synthesis. FUT3, FUT4, FUT5, FUT6, FUT7 and FUT9 are alpha 1-3/4 Fucosyltransferases and are responsible for Lewis structure generation. FUT5 has high homology with FUT3 and FUT6 due to gene duplication. FUT7 is exclusively responsible for biosynthesis of sialyl Lewis X epitope in leukocytes and high endothelial venule cells (2). FUT8 is an alpha 1-6 Fucosyltransferase that adds a fucose to the chitobiose core of N-glycans (3). Predicted as type II transmembrane proteins and Golgi enzymes, some of the Fucosyltransferases can also be found in plasma. R&D Systems recombinant human FUTs correspond to the luminal domains. The activity of this enzyme has been measured using a phosphatase-coupled method (4).
Recombinant Human Fucosyltransferase 5/FUT5 Protein, CF
R&D Systems | Catalog # 4949-GT
FUT5 has been formulated so that it can be used in the cell surface glycoengineering of living cells, and does not affect cell viability or native phenotype apart from the intended impact on cell glycobiology.
Loading...
Key Product Details
- R&D Systems NS0-derived Recombinant Human Fucosyltransferase 5/FUT5 Protein (4949-GT)
- 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
Structure / Form
Disulfide-linked homodimer
Applications
Enzyme Activity
Loading...
Learn more about Fluorescent Glycan Labeling and Detection
Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Fucosyltransferase 5/FUT5 protein
Arg35-Thr374, with an N-terminal 6-His tag
Arg35-Thr374, with an N-terminal 6-His tag
Purity
>80%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
His
Predicted Molecular Mass
40 kDa
SDS-PAGE
58 kDa, reducing conditions
Activity
Measured by its ability to transfer fucose from GDP-fucose to N-Acetyllactosamine
The specific activity is >35 pmol/min/µg, as measured under the described conditions.
The specific activity is >35 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
4949-GT
| 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.
|
Background: Fucosyltransferase 5/FUT5
References
- Becker, D.J. et al. (2003) Glycobiology 13:41R.
- Blander, J. M. et al. (1999) J. Immunol. 163:3746.
- Lee, S.H. et al. (2006) J. Biochem. 139:391.
- Wu, Z.L. et al. (2010) Glycobiology doi: 10.1093/glycob/cwq187.
Alternate Names
FUC-TV, FUT5
Entrez Gene IDs
2527 (Human)
Gene Symbol
FUT5
UniProt
Additional Fucosyltransferase 5/FUT5 Products
Product Documents for Recombinant Human Fucosyltransferase 5/FUT5 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Fucosyltransferase 5/FUT5 Protein, CF
For research use only
Related Research Areas
Customer Reviews for Recombinant Human Fucosyltransferase 5/FUT5 Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human Fucosyltransferase 5/FUT5 Protein, CF and earn rewards!
Have you used Recombinant Human Fucosyltransferase 5/FUT5 Protein, CF?
Submit a review and receive an Amazon gift card!
$25/€18/£15/$25CAN/¥2500 Yen for a review with an image
$10/€7/£6/$10CAN/¥1110 Yen for a review without an image
Submit a review
Protocols
View specific protocols for Recombinant Human Fucosyltransferase 5/FUT5 Protein, CF (4949-GT):
Materials
- Assay Buffer: 25 mM Tris, 5 mM MnCl2 (supplied in kit), pH 7.5
- Recombinant Human Fucosyltransferase 5/FUT5 FUT5 (rhFUT5) (Catalog # 4949-GT)
- Donor Substrate: GDP-Fucose (Sigma, Catalog # G4401), 1.6 mM stock in deionized water
- Acceptor Substrate: Lactosamine (V-Labs, Catalog # GN204), 50 mM in deionized water
- Glycosyltransferase Activity Kit (Catalog # EA001)
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute GDP-Fucose to 240 μM in Assay Buffer.
- Dilute Lactosamine to 2.4 mM in Assay Buffer.
- Dilute Coupling Phosphatase 1 to 12 μg/mL in Assay Buffer.
- Prepare reaction mixture by combining equal volumes of diluted GDP-Fucose, Lactosamine, and Coupling Phosphatase 1.
- Dilute rhFUT5 to 20 µg/mL in Assay Buffer.
- Dilute 1 mM Phosphate Standard by adding 40 µL of the 1 mM Phosphate Standard to 360 µL of Assay Buffer for a 100 µM stock.
- Prepare standard curve by performing seven one-half serial dilutions of the 100 µM Phosphate stock in Assay Buffer. The standard curve has a range of 0.039 to 2.5 nmol per well.
- Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of Assay Buffer.
- Load 25 µL of the 20 µg/mL rhFUT5 into the plate. Include a Blank Control containing 25 µL of Assay Buffer.
- Add 25 µL of reaction mixture (step 4) to the wells, excluding the standard curve and curve blank.
- Cover the plate with parafilm or a plate sealer and incubate at 37 ºC for 20 minutes.
- Add 30 µL of the Malachite Green Reagent A to all wells. Tap to mix briefly.
- Add 100 µL of deionized water to all wells. Tap to mix briefly.
- Add 30 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
- Read plate at 620 nm (absorbance) in endpoint mode.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Phosphate released* (nmol) x (1000 pmol/nmol) |
| Incubation time (min) x amount of enzyme (µg) |
*Derived from the phosphate standard curve using linear or 4-parameter fitting and adjusted for Blank Control.
Per Reaction:
- rhFUT5: 0.5 µg
- Coupling Phosphatase 1: 0.1 µg
- Lactosamine: 400 µM
- GDP-Fucose: 40 µM
Loading...