Recombinant Human ST6GAL2 Protein, CF
R&D Systems | Catalog # 8330-GT
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Key Product Details
- R&D Systems CHO-derived Recombinant Human ST6GAL2 Protein (8330-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 ST6 Gal Sialyltransferase 2/ST6GAL2 protein
Glu243-Ser529, with a C-terminal 6-His tag
Glu243-Ser529, with a C-terminal 6-His tag
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Endotoxin Level
<0.1 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Glu243
Predicted Molecular Mass
33 kDa
SDS-PAGE
37-43 kDa, reducing conditions
Activity
Measured by its ability to transfer Neu5Ac from CMP-Neu5Ac to fetuin of fetal calf serum.
The specific activity is >0.45 pmol/min/μg, as measured under the described conditions.
The specific activity is >0.45 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
8330-GT
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol. |
| 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: ST6 Gal Sialyltransferase 2/ST6GAL2
References
- Varki, A. et al. (1999) Essentials of Glycobiology, ColdSpring Harbor Laboratory Press, pp195.
- Takashima, S. et al. (2002) J. Biol. Chem. 277:45719.
- Lehoux, S. et al. (2009) Glycoconj. J. 27:99.
- Takashima, S. et al. (2003) J. Biochem. 134:287.
- Laporte, B. et al. (2009) Glycobiology 19:1082.
Long Name
ST6 beta-Galactoside alpha-2,6-Sialyltransferase 2
Alternate Names
Alpha 2,6-ST 2, KIAA1877, Sialyltransferase 2, SIAT2
Gene Symbol
ST6GAL2
UniProt
Additional ST6 Gal Sialyltransferase 2/ST6GAL2 Products
Product Documents for Recombinant Human ST6GAL2 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 ST6GAL2 Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Human ST6GAL2 Protein, CF (8330-GT):
Materials
- Sialyltransferase Activity Kit (Catalog # EA002)
- 10X Assay Buffer (supplied in kit): 250 mM Tris, pH 7.5
- MnCl2 (supplied in kit): 100 mM
- Recombinant Human ST6 Gal Sialyltransferase 2/ST6GAL2 (rhST6GAL2) (Catalog # 8330-GT)
- Recombinant C. perfringens Neuraminidase (rCpNA) (Catalog # 5080-NM)
- CMP-Sialic Acid (Sigma, Catalog # C8271), 10 mM stock in deionized water
- Fetuin (Sigma, Catalog # F3385), 50 mg/mL in deionized water
- 96-well Clear Plate (R&D Systems, Catalog # DY990)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Prepare 1X Assay Buffer containing 10 mM MnCl2 by combining 10X stocks and diluting 10-fold with deionized water.
- Dilute 1 mM Phosphate Standard provided by the Sialyltransferase Activity Kit by adding 40 µL of the 1 mM Phosphate Standard to 360 µL of 1X Assay Buffer for a 100 µM stock. This is the first point of the standard curve.
- Complete the standard curve by performing six one-half serial dilutions of the 100 µM Phosphate stock using 1X Assay Buffer. The standard curve has a range of 0.078 to 5 nmol per well.
- Prepare reaction mixture containing 0.4 mM CMP-Sialic Acid, 10 mg/mL Fetuin, 8 µg/mL rCpNA and 4 µg/mL Coupling Phosphatase 2 in 1X Assay Buffer.
- Dilute rhST6Gal2 to 80 µg/mL in 1X Assay Buffer.
- Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of 1X Assay Buffer.
- Load 25 µL of the 80 µg/mL rhST6Gal2 into the plate. Include a control containing 25 µL of 1X Assay Buffer.
- Add 25 µL of reaction mixture to the wells, excluding the standard curve.
- Seal plate and incubate at room temperature overnight.
- 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 fitting and adjusted for Control.
Per Reaction:
- rhST6Gal2: 2 µg
- Coupling Phosphatase 2: 0.1 µg
- rCpNA: 0.2 µg
- Fetuin: 0.25 mg
- CMP-Sialic Acid: 200 µM
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