Recombinant Human ST3GAL1 Protein, CF

R&D Systems | Catalog # 6905-GT

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human ST3GAL1 Protein (6905-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

Q11201

Applications

Enzyme Activity
View all ST3 beta-Gal alpha-2,3-Sialyltransferase 1/ST3GAL1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human ST3 beta-Gal alpha-2,3-Sialyltransferase 1/ST3GAL1 protein
Arg57-Arg340, with an N-terminal HA (YPYDVPDYA) tag

Purity

>85%, 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.

N-terminal Sequence Analysis

Tyr (of HA tag)

Predicted Molecular Mass

33.4 kDa

SDS-PAGE

35-45 kDa, reducing conditions

Activity

Measured by its ability to transfer Neu5Ac from CMP-Neu5Ac to bovine B1-3 galactosyl-N-acetyl galactosamine.
The specific activity is >1000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6905-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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: ST3 beta-Gal alpha-2,3-Sialyltransferase 1/ST3GAL1

Sialyltransferases add sialic acid to glycoproteins or glycosphingolipids and play important roles in many biological processes including immune recognition, pathogen infection, and cell adhesion (1). ST3GAL1 is a type II membrane protein localized in the trans‑Golgi network that transfers sialic acid to galactose residues of core 1 O‑glycans (3Gal beta 1‑3GalNAc) to produce sialyl‑T antigen (NeuAc alpha 2‑3Gal beta 1‑3GalNAc) (2, 3). ST3GAL1 is ubiquitously expressed in most tissues, with highest expression in the placenta, liver and skeletal muscle (4). Over‑expression of ST3GAL1 may be involved in the tumorigenesis of both breast cancer (5, 6) and bladder cancer (7). The enzyme activity of the recombinant human ST3GAL1 was determined using a phosphatase‑coupled glycosyltransferase assay (8).

References

  1. Varki, A. (1999) Glycobiology 2:25.
  2. Jeanneau, C. et al. (2004) J. Biol. Chem. 279:13461.
  3. Dalziel, M. et al. (2001) J. Biol. Chem. 276:11007.
  4. Kitagawa, H. and Paulson, J.C. (1994) J. Biol. Chem. 269:17872.
  5. Picco, G. et al. (2010) Glycobiology 20:1241.
  6. Burchell, J. et al. (1999) Glycobiology 9:1307.
  7. Videira, P.A. et al. (2009) BMC Cancer 9:357.
  8. Wu, Z.L. et al. (2011) Glycobiology 21:727.

Long Name

ST3 beta-Galactoside alpha-2,3-Sialyltransferase 1

Alternate Names

Gal-NAc6S, SIAT4A, SIATFL, ST3GalA, ST3O

Entrez Gene IDs

6482 (Human); 20442 (Mouse); 362924 (Rat)

Gene Symbol

ST3GAL1

UniProt

Q11201

Additional ST3 beta-Gal alpha-2,3-Sialyltransferase 1/ST3GAL1 Products

Product Documents for Recombinant Human ST3GAL1 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.

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Product Specific Notices for Recombinant Human ST3GAL1 Protein, CF

For research use only

Related Research Areas

Glycobiology-related Enzymes

Citations for Recombinant Human ST3GAL1 Protein, CF

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Protocols

View specific protocols for Recombinant Human ST3GAL1 Protein, CF (6905-GT):

Materials
  • Sialyltransferase Activity Kit (Catalog # EA002)
  • 10X Assay Buffer: 250 mM Tris, pH 7.5
  • MnCl2 (supplied in kit): 100 mM
  • Recombinant Human beta -Gal alpha -2,3-Sialyltransferase/ST3GAL1 (rhST3GAL1) (Catalog # 6905-GT)
  • CMP-Neu5Ac (Sigma, Catalog # C8271), 10 mM stock in deionized water
  • beta 1-3 Galactosyl-N-acetyl galactosamine (V-labs, Catalog # GN213), 50 mM stock in deionized water
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Prepare 1X Assay Buffer containing 10 mM MnCl2 by combining equal volumes of 10X Assay Buffer and 100 mM MnCl2 and diluting 5-fold with deionized water.
  2. Dilute Phosphate Standard 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.
  3. 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 nmoles per well.
  4. Prepare reaction mixture containing 0.4 mM CMP-Neu5Ac, 1 mM beta 1-3 Galactosyl-N-acetyl galactosamine, 4 μg/mL Coupling Phosphatase 2 in 1X Assay Buffer.
  5. Dilute rhST3GAL1 to 1 µg/mL in Assay Buffer.
  6. Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of 1X Assay Buffer.
  7. Load 25 µL of the 1 µg/mL rhST3GAL1 into the plate. Include a Control containing 25 µL of 1X Assay Buffer.
  8. Add 25 µL of reaction mixture to the wells, excluding the standard curve.
  9. Seal plate and incubate at 37 °C for 20 minutes.
  10. Add 30 µL of the Malachite Green Reagent A to all wells. Mix briefly.
  11. Add 100 µL of deionized water to all wells. Mix briefly.
  12. Add 30 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
  13. Read plate at 620 nm (absorbance) in endpoint mode.
  14. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted 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:

  • rhST3GAL1: 0.025 μg
  • Coupling Phosphatase 2: 0.1 μg
  • CMP-Neu5Ac: 200 µM
  • beta 1-3 Galactosyl-N-acetyl galactosamine: 0.5 mM

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