Recombinant Human Fucosyltransferase 2/FUT2 Protein, CF

Catalog # Availability Size / Price Qty
7770-GT-020
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Human Fucosyltransferase 2/FUT2 Protein, CF Summary

Learn more about Fluorescent Glycan Labeling and Detection

Product Specifications

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.
Activity
Measured by its ability to transfer fucose from GDP-fucose to lactose. The specific activity is >2,500 pmol/min/μg, as measured under the described conditions.
Source
Human embryonic kidney cell, HEK293-derived human Fucosyltransferase 2/FUT2 protein
Ile35-His343, with an C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Ile35
Predicted Molecular Mass
36 kDa
SDS-PAGE
40-50 kDa, reducing conditions

Product Datasheets

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7770-GT

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.

7770-GT

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, DTT and Glycerol.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 25 mM Tris, 150 mM NaCl, 10 mM MnCl2, 10 mM CaCl2, pH 7.5
  • Recombinant Human FUT2 (rhFUT2) (Catalog # 7770-GT)
  • GDP-Fucose (Sigma, Catalog # G4401), 1.6 mM stock in deionized water
  • alpha -lactose (Sigma, Catalog # L2643), 0.3 M stock 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
  1. Dilute 1 mM Phosphate Standard provided by the Glycosyltransferase Kit to 100 µM by adding 40 µL of standard to 360 µL of Assay Buffer. This is the first point of the standard curve.
  2. Perform six additional one-half serial dilutions in Assay Buffer. The standard curve has a range of 0.078 to 5 nmol per well.
  3. Prepare reaction mixture containg 100 mM alpha -lactose, 200 µM GDP-Fucose, and 4 µg/mL Coupling Phosphatase I in Assay Buffer.
  4. Dilute rhFUT2 to 1.0 µg/mL in Assay Buffer.
  5. Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of Assay Buffer.
  6. Load 25 µL of the 1 µg/mL rhFUT2 into the plate. Include a Substrate Blank containing 25 µL of Assay Buffer.
  7. Add 25 µL of reaction mixture (step 3) to the wells, excluding the standard curve and curve blank.
  8. Cover the plate with a plate sealer and incubate at 37 °C for 20 minutes.
  9. Add 30 µL of the Malachite Green Reagent A to all wells. Mix briefly.
  10. Add 100 µL of deionized water to all wells. Mix briefly.
  11. Add 30 µL of the Malachite Green Reagent B to all wells. Mix and inubate for 20 minutes at room temperature.
  12. Read plate at 620 nm (absorbance) in endpoint mode.
  13. 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 Control.

Per Reaction:
  • rhFUT-2: 0.025 µg
  • Coupling Phosphatase I: 0.1 µg
  • alpha -Lactose: 50 mM
  • GDP-Fucose: 100 µM
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: Fucosyltransferase 2/FUT2

Glycans are frequently fucosylated at terminal sites. Therefore, fucose is often part of a sugar epitope with important biological function. Well‑known fucose‑containing glycans include Lewis and ABO blood group antigens. Lewis epitopes are key elements involved in leukocyte homing and the extravasation process, and thus are important for lymphocyte maturation and natural defense functions. Fucose‑containing glycans also play critical roles in cell signaling and development (1). More than 10 fucosyltransferases have been cloned, and most are Golgi‑resident type II membrane proteins (2). FUT3, FUT4, FUT5, FUT6, FUT7, and FUT9 are alpha 1‑3 or  alpha 1‑4 fucosyltransferases and are responsible for Lewis antigen generation (3, 4, 5). FUT8 is the only alpha 1‑6 fucosyltransferase that adds a fucose to the chitobiose core of N‑glycans (6). FUT1 and FUT2 are galactoside alpha 1‑2 fucosyltransferases that generate H‑antigen, the precursor for ABO blood‑group antigen synthesis. In particular, FUT2 involves in generating soluble ABO antigen in saliva; therefore it is also called secretor blood group alpha 1‑2 fucosyltransferase (7). Absence of functional FUT2 is related to Crohn’s disease (8). The activity of this enzyme has been measured with a phosphatase‑coupled method (9).

References
  1. Jafar-Nejad, H. et al. (2010) Glycobiology 20:931.
  2. Becker, D.J. et al. (2003) Glycobiology 13:41R.
  3. Blander, J. M. et al. (1999) J. Immunol. 163:3746.
  4. Natsuka, S. et al. (1994) J. Biol. Chem. 269:16789.
  5. Sasaki, K. et al. (1994) J. Biol. Chem. 269:14730.
  6. Lee, S.H. et al. (2006) J. Biochem. 139:391.
  7. Kelly, R.J. et al. (1995) J. Biol. Chem. 270:4640.
  8. McGovern, D.P. et al. (2010) Hum. Mol. Genet. 19:3468.
  9. Wu, Z.L. et al. (2011) Glycobiology 21:727.
Entrez Gene IDs
2524 (Human); 14344 (Mouse); 58924 (Rat)
Alternate Names
alpha (1,2) fucosyltransferase; Alpha(1,2)FT 2; alpha(1,2)FT2; B12QTL1; EC 2.4.1.69; fucosyltransferase 2 (secretor status included); Fucosyltransferase 2; FUT2; galactoside 2-alpha-L-fucosyltransferase 2; GDP-L-fucose:beta-D-galactoside 2-alpha-L-fucosyltransferase 2; Se; Se2; SEC2; SEC2SE; Secretor blood group alpha-2-fucosyltransferase; Secretor factor; SEJ

Product Specific Notices

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

Citation for Recombinant Human Fucosyltransferase 2/FUT2 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.

1 Citation: Showing 1 - 1

  1. Towards Mapping of the Human Brain N-Glycome with Standardized Graphitic Carbon Chromatography
    Authors: J Helm, L Hirtler, F Altmann
    Biomolecules, 2022;12(1):.
    Species: Human
    Sample Types: Small Molecule
    Applications: Bioassay

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