beta 4GalT1 is one of seven beta 1,4 galactosyltransferases that transfer galactose in a beta 1,4 linkage to acceptor sugars including GlcNAc, and Glc, and Xyl. By sequence similarity, the beta 4GalTs form four groups: beta 4GalT1 and beta 4GalT2, beta 4GalT3 and beta 4GalT4, beta 4GalT5 and beta 4GalT6, and beta 4GalT7 (1). beta 4GalT1 is unique among the seven enzymes because it can be expressed either as membrane associated form or secreted form (2). The secreted form is restricted to lactating mammary tissues where the enzyme forms a heterodimer with alpha -lactalbumin to catalyze the synthesis of lactose (3). The membrane form can reside either in the Golgi apparatus, where it adds galactose to N-acetylglucosamine residues, or on cell surface, where it functions as a recognition molecule during a variety of cell to cell and cell to matrix interactions, by binding to specific oligosaccharide ligands on opposing cells or in the extracellular matrix (4). The two enzymatic forms result from alternate transcription initiation sites and post-translational processing (5). Defects in beta 4GalT1 are the cause of congenital disorder of glycosylation type 2D (CDG2D) (6). The activity of this enzyme has been measured with a phosphatase-coupled method (7).
Recombinant Human B4GalT1 Protein, CF
R&D Systems | Catalog # 3609-GT
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Key Product Details
- R&D Systems NS0-derived Recombinant Human B4GalT1 Protein (3609-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
Applications
Enzyme Activity
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Learn more about Fluorescent Glycan Labeling and Detection
Product Specifications
Source
Mouse myeloma cell line, NS0-derived human beta-1,4-Galactosyltransferase 1/B4GalT1 protein
Gly44-Ser398, with an N-terminal 6-His tag
Gly44-Ser398, with an N-terminal 6-His tag
Purity
>95%, 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
45-55 kDa
Activity
Measured by its ability to transfer galactose from UDP-galactose to N-Acetyl-alpha -D-glucosamine.
The specific activity is >2,000 pmol/min/μg, as measured under the described conditions.
The specific activity is >2,000 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
3609-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: beta-1,4-Galactosyltransferase 1/B4GalT1
References
- Amado, M. et al. (1999) Biochim. Biophys. Acta. 1473:35.
- Yamaguchi, N and Fukuda, M.N. (1995) J. Biol. Chem. 270:12170.
- Appert, H.E. et al. (1986) Biochem. Biophys. Res. Commun. 138:224.
- Lopez, L.C. et al. (1991) J. Biol. Chem. 266:15984.
- Mengle-Gaw, L. et al. (1991) Biochem. Biophys. Res. Commun. 176:1269.
- Hansske, B. et al. (2002) J. Clin. Invest. 109:725.
- Wu, Z.L. et al. (2011) Glycobiology 21:727.
Long Name
UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase, Polypeptide 1
Alternate Names
GGTB2
Gene Symbol
B4GALT1
UniProt
Additional beta-1,4-Galactosyltransferase 1/B4GalT1 Products
Product Documents for Recombinant Human B4GalT1 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 B4GalT1 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human B4GalT1 Protein, CF
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Protocols
View specific protocols for Recombinant Human B4GalT1 Protein, CF (3609-GT):
Materials
- Assay Buffer: 25 mM Tris, 150 mM NaCl, 10 mM MgCl2, 10 mM MnCl2 (supplied in kit), pH 7.5
- Recombinant Human beta -1,4‑Galactosyltransferase 1/B4GalT1 (rhB4GalT1) (Catalog # 3609-GT)
- Donor Substrate: UDP-Galactose (Sigma, Catalog # U4500), 10 mM stock in deionized water
- Acceptor Substrate: N-Acetyl-alpha -D-glucosamine (GlcNAc) (EMD, Catalog # 1079), 1 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
- Dilute UDP-Galactose to 0.5 mM in Assay Buffer.
- Dilute Coupling Phosphatase 1 to 20 µg/mL in Assay Buffer.
- Dilute GlcNAc to 50 mM in Assay Buffer.
- Prepare Reaction Mixture by combining equal volumes of 0.5 mM UDP-GalNAc, 20 µg/mL Coupling Phosphatase 1, and 50 mM GlcNAc.
- Dilute rhB4GalT1 to 0.75 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. This is the first point of the standard curve.
- Continue standard curve by performing six one-half serial dilutions of the 100 µM Phosphate stock in Assay Buffer. The standard curve has a range of 0.078 to 5.0 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 20 µL of the 0.75 µg/mL rhB4GalT1 into the plate. Include a Control containing 20 µL of Assay Buffer.
- Start the reaction by adding 30 µL of Reaction Mixture to the wells, excluding the standard curve and curve blank.
- Seal plate and incubate at room temperature for 15 minutes.
- Add 30 µL of the Malachite Green Reagent A to all wells. Mix briefly.
- Add 100 µL of deionized water to all wells. 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 Control.
Per Well:
- rhB4GalT1: 0.015 µg
- Coupling Phosphatase 1: 0.2 µg
- UDP-Galactose: 5 nmol
- GlcNAc: 500 nmol
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