Polysialic acid (PSA), a
glycan abundant on the neural cell adhesion molecule (NCAM) during embryonic
development, negatively modulates the adhesive properties of NCAM (1).
Following birth, PSA expression decreases promptly and becomes restricted to
the hippocampus, hypothalamus, and olfactory bulb-areas of the brain that
require continuous cell migration and synaptic plasticity (2). Expression of
PSA in cancer cells has been suggested to increase tumor invasiveness and
promote tumor growth (3). However; the mouse St8sia6 only has alpha -2,8 sialyltransferase activity toward both glycolipids and glycoproteins that has the NeuAc-alpha -2,3(6)-beta -D-Gal sequence at the nonreducing ends of their carbohydrate groups, and forms NeuAc-alpha -2,8-NeuAc structures, but not oligosialic or polysialic acid structures (4). Like most glycosyltransferases, ST8SIA6 may be a Golgi‑resident type II membrane protein. It is expressed in various tissues at low levels but is highly expressed in breast cancer and Dami megakaryocyte cell lines (5). The activity of this enzyme has been measured with a phosphatase-coupled method (6).
Recombinant Human ST8SIA6 Protein, CF
R&D Systems | Catalog # 9587-GT
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Key Product Details
- R&D Systems CHO-derived Recombinant Human ST8SIA6 Protein (9587-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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Learn more about Fluorescent Glycan Labeling and Detection
Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived human ST8 alpha-2,8-Sialyltransferase 6/ST8SIA6 protein
Asp28-Ala398, with an N-terminal 6-His tag
Asp28-Ala398, with an N-terminal 6-His tag
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
His
Predicted Molecular Mass
43 kDa
SDS-PAGE
62-73 kDa, reducing conditions
Activity
Measured by its ability to transfer Neu5Ac from CMP-Neu5Ac to fetuin of fetal calf serum.
The specific activity is >70 pmol/min/μg, as measured under the described conditions.
The specific activity is >70 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
9587-GT
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| 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.
|
Background: ST8 alpha-2,8-Sialyltransferase 6/ST8SIA6
References
- Scheidegger, E.P. et al. (1995) J. Biol. Chem. 270:22685.
- Rutishauser, U. (2008) Nat. Rev. Neurosci. 9:26.
- Seidenfaden, R. et al. (2003) Mol.Cell. Biol. 23:5908.
- Takashima, S. et al. (2002) J. Biol. Chem. 277:24030.
- Teintenier-Lelievre, M. et al. (2005) Biochem. J. 392: 665.
- Wu, Z.L. et al. (2011) Glycobiology 21:727.
Long Name
ST8 alpha-N-acetyl-neuraminide alpha-2,8-sialyltransferase 6
Alternate Names
SIA8F, Sialyltransferase 8F, SIAT8F, ST8 alpha-2,8-Sialyltransferase 6, ST8Sia VI, ST8SIA-VI
Gene Symbol
ST8SIA6
UniProt
Additional ST8 alpha-2,8-Sialyltransferase 6/ST8SIA6 Products
Product Documents for Recombinant Human ST8SIA6 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 ST8SIA6 Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Human ST8SIA6 Protein, CF (9587-GT):
Materials
- Sialyltransferase Activity Kit (Catalog # EA002)
- Assay Buffer: 50 mM MES, 10 mM MnCl2, pH 6.5
- Recombinant Human ST8 alpha-2,8-Sialyltransferase (rhST8SIA6) (Catalog # 9587-GT)
- CMP-Neu5Ac (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 (Catalog # DY990)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- 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 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 Assay Buffer. The standard curve has a range of 0.078 to 5 nmol per well.
- Prepare reaction mixture containing 0.5 mM CMP-Neu5Ac, 20 mg/mL Fetuin, and 4 µg/mL coupling phosphatase 2 (supplied in kit) in Assay Buffer.
- Dilute rhST8SIA6 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 the 20 µg/mL rhST8SIA6 into empty wells of the same plate as the curve. Include a control containing 25 µL of Assay Buffer.
- Add 25 µL of reaction mixture to the wells, excluding the standard curve.
- Incubate sealed plate 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:
- rhST8SIA6: 0.5 µg
- Coupling Phosphatase 2: 0.1 µg
- CMP-Neu5Ac: 250 µM
- Fetuin: 500 µg