Recombinant Human ST6GALNAC5 Protein, CF

R&D Systems | Catalog # 6715-GT

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human ST6GALNAC5 Protein (6715-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

Q9BVH7

Applications

Enzyme Activity
View all ST6 GalNAc alpha-2,6-sialyltransferaseV/ST6GALNAC5 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human ST6 GalNAc alpha-2,6-sialyltransferaseV/ST6GALNAC5 protein
Gly30-Phe336, with an N-terminal 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

His

Predicted Molecular Mass

36 kDa

SDS-PAGE

40-55 kDa, reducing conditions

Activity

Measured by its ability to transfer Neu5Ac from CMP-Neu5Ac to fetuin of fetal calf serum.
The specific activity is >40 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6715-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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: ST6 GalNAc alpha-2,6-sialyltransferaseV/ST6GALNAC5

Gangliosides are acidic glycosphingolipids that contain one or more sialic acid residues (1). They are abundant in the nervous system, where they play crucial modulatory roles in cellular recognition, interaction, adhesion, and signal transduction, particularly during early developmental stages. The expression of gangliosides in the nervous system is developmentally regulated through various sialyltransferases (2). ST6GALNAC5 is a sialyltransferase involved in the biosynthesis of ganglioside GD1a (NeuAc alpha 2,3Gal beta 1,3GalNAc beta 1,4(NeuAc alpha 2,3)Gal beta 1,4Glc beta 1-Cer) from GM1b (NeuAc alpha 2,3Gal beta 1,3GalNAc beta 1,4Gal beta 1,4Glc beta 1-Cer), and its expression is restricted to the brain (3, 4). ST6GALNAC5 has been identified as a key player in the metastasis of breast cancer cells to the brain by potentially enabling the cancer cells to cross the blood-brain barrier (5, 6). The recombinant ST6GALNAC5 was active on fetuin from fetal calf serum when assayed using a phosphatase-coupled method (7) suggesting the substrate specificity of ST6GALNAC5 may require further characterization.

References

  1. Kolter, T. et al. (2002) J. Biol. Chem. 277:25859.
  2. Yu, R.K. et al. (2008) Glycoscience DOI: 10.1007/978-3-540-30429-6_41.
  3. Okajima, T. et al. (1999) J. Biol. Chem.274:30557.
  4. Harduin-Lepers, A. et al. (2005) Glycobiology 15:805.
  5. Bos, P.D. et al. (2009) Nature 459:1005.
  6. Arshad, F. et al. (2011). Patholog. Res. Int. Doi: 10.4061/2011/920509.
  7. Wu, Z.L. et al. (2010) Glycobiology doi: 10.1093/glycob/cwq187.

Long Name

ST6 alpha-N-acetylGalactosaminide alpha-2,6-sialyltransferase 5

Alternate Names

GD1 alpha synthase, SIAT7E, ST6GalNAcV

Entrez Gene IDs

81849 (Human); 26938 (Mouse)

Gene Symbol

ST6GALNAC5

UniProt

Q9BVH7

Additional ST6 GalNAc alpha-2,6-sialyltransferaseV/ST6GALNAC5 Products

Product Documents for Recombinant Human ST6GALNAC5 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 ST6GALNAC5 Protein, CF

For research use only

Related Research Areas

Glycobiology-related Enzymes

Citations for Recombinant Human ST6GALNAC5 Protein, CF

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Protocols

View specific protocols for Recombinant Human ST6GALNAC5 Protein, CF (6715-GT):

Materials
  • Sialyltransferase Activity Kit (Catalog # EA002)
  • Assay Buffer: 25 mM MES, 20 mM MgCl2, pH 6.5
  • Recombinant Human GalNAc alpha -2,6-sialyltransferase V/ST6GALNA (rhST6GALNAC5) (Catalog # 6715-GT)
  • CMP-Neu5Ac (Sigma, Catalog # C8271), 10 mM stock in deionized water
  • Fetuin (Sigma, Catalog # F3385), 50 mg/mL stock in deionized water
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent

  1. 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.
  2. 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 nmoles per well.
  3. Dilute CMP-Neu5Ac to 1.5 mM in Assay Buffer.
  4. Dilute Fetuin to 30 mg/mL in Assay Buffer.
  5. Dilute Coupling Phosphatase 2 to 12 μg/mL in Assay Buffer.
  6. Prepare reaction mixture by combining equal volumes of diluted CMP-Neu5Ac, Fetuin, and Coupling Phosphatase 2.
  7. Dilute rhST6GALNAC5 to 10 µg/mL in Assay Buffer.
  8. Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of Assay Buffer.
  9. Load 25 µL of the 10 µg/mL rhST6GALNAC5 into the plate. Include a Control containing 25 µL of Assay Buffer.
  10. Add 25 µL of reaction mixture to the wells, excluding the standard curve.
  11. Cover the plate with parafilm or a plate sealer and incubate at 37 °C for 20 minutes.
  12. Add 30 µL of the Malachite Green Reagent A to all wells. Mix briefly.
  13. Add 100 µL of deionized water to all wells. Mix briefly.
  14. Add 30 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
  15. Read plate at 620 nm (absorbance) in endpoint mode.
  16. 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:

  • rhST6GALNAC5: 0.25 μg
  • Coupling Phosphatase 2: 100 ng
  • CMP-Neu5Ac: 250 µM
  • Fetuin: 250 μg

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