Recombinant Human B4GAT1 His-tag Protein, CF
R&D Systems | Catalog # 6664-GT
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Key Product Details
- R&D Systems CHO-derived Recombinant Human B4GAT1 His-tag Protein (6664-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
CHO
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived human beta-1,4-Glucuronyltransferase 1/B4GAT1 protein
His37-Cys415, with N-terminal 6-His tag
His37-Cys415, with N-terminal 6-His tag
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Predicted Molecular Mass
43.8 kDa
SDS-PAGE
42-53 kDa under reducing conditions
Activity
Measured by its ability to transfer GlcA from UDP-GlcA to Xylose.
The specific actiivity is >55 pmol/min/μg, as measured under the described conditions.
The specific actiivity is >55 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
6664-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-Glucuronyltransferase 1/B4GAT1
References
- Sasaki, K. et al. (1997) PNAS 94:14294.
- Willer, T. et al. (2014) eLife; 3: e03941.
- Barresi, R. and Campbell, K.P. (2006) J. Cell Sci. 119:199.
- Wu, Z.L. et al. (2011) Glycobiology 21:727.
Long Name
beta-1,4-Glucuronyltransferase 1
Alternate Names
B3GNT1, iGAT, iGNT, MDDGA13
Gene Symbol
B4GAT1
UniProt
Additional beta-1,4-Glucuronyltransferase 1/B4GAT1 Products
Product Documents for Recombinant Human B4GAT1 His-tag 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 B4GAT1 His-tag Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Human B4GAT1 His-tag Protein, CF (6664-GT):
Materials
- Glycosyltransferase Activity Kit (Catalog # EA001)
- Assay Buffer: 25 mM Tris, 10 mM CaCl2, 10 mM MnCl2 (supplied in kit), pH 7.5
- Recombinant Human beta -1,4-Glucuronyltransferase 1/B4GAT1 His-tag (rhB4GAT1) (Catalog # 6664-GT)
- Donor Substrate: UDP-GlcA (Sigma, Catalog # U5625), 10 mM stock in deionized water
- Acceptor Substrate: Xylose (V-lab, Catalog # BX53), 100 mM stock in deionized water
- 96-well Clear Plate (Catalog # DY990)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute 1 mM Phosphate Standard provided by the Glycosyltransferase Kit 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.
- Prepare 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 nmol per well.
- Prepare reaction mixture containing 0.8 mM UDP-GlcA, 40 mM Xylose, and 4 µg/mL Coupling Phosphatase I in Assay Buffer.
- Dilute rhB4GAT1 to 20 µg/mL in Assay Buffer.
- 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 20 µg/mL rhB4GAT1 into the plate. Include a Control containing 25 µL of Assay Buffer.
- Add 25 µL of reaction mixture to all wells, excluding the standard curve and curve blank.
- Seal plate and incubate at 37 °C for 20 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 Reaction:
- rhB4GAT1: 0.5 µg
- Coupling Phosphatase I: 0.1 µg
- Xylose: 20 mM
- UDP-GlcA: 0.4 mM
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