Recombinant Human ST6GAL1 (aa 44-406) Protein, CF

R&D Systems | Catalog # 7620-GT

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems NS0-derived Recombinant Human ST6GAL1 (aa 44-406) Protein (7620-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

P15907

Applications

Enzyme Activity
View all ST6 Gal Sialyltransferase 1/ST6GAL1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human ST6 Gal Sialyltransferase 1/ST6GAL1 protein
Glu44-Cys406 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

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

N-terminal Sequence Analysis

His

Predicted Molecular Mass

42 kDa

SDS-PAGE

40-50 kDa, reducing conditions

Activity

Measured by its ability to transfer Neu5Ac from CMP-Neu5Ac to N-Acetyllactosamine.
The specific activity is >150 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 4 using 7620-GT in the following applications:

Formulation, Preparation, and Storage

7620-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 Gal Sialyltransferase 1/ST6GAL1

Sialic acid molecules attached to glycoproteins or glycosphingo lipids play important roles in various biological processes such as immune recognition, pathogen infection, and cell adhesion (1).Sialyltransferases are key enzymes that regulate the cellular levels of sialic acid containing molecules. Beta-galactosamide alpha‑2,6‑sialyltransferase 1 encoded by the ST6GAL1 gene is a type II membrane protein localized in the trans-Golgi network and catalyzes 2,6-sialylation of Gal beta 1,4‑GlcNAc structures on N-glycans (2). The enzyme is involved in the generation of the cell surface carbohydrate determinants and differentiation antigens HB6, CD75, and CD76 (3). ST6GAL1 is highly expressed in the liver and also expressed in most other tissues to some extent (4). ST6GAL1 deficiency causes abnormalities in B‑cell immunoreactivity (5). The expression and activity of ST6GAL1 are associated with tumor metastasis in breast (6) and colon (7) cancers. The majority of ST6GAL1 in the liver is cleaved and secreted into the serum (8) and may be used as a biomarker for hepatitis diseases (9). Using a phosphatase-coupled sialyltransferase assay, the Km for the donor substrate CMP-NeuAc was determined to be around 0.5 mM (10).

References

  1. Varki, A. et al. (1999) Essentials of Glycobiology, Cold Spring Harbor Laboratory Press, pp195.
  2. Weinstein, J. et al. (1987) J. Biol. Chem. 262:17735.
  3. Bast, B.J. et al. (1992) J. Cell Biol. 116:423.
  4. Kitagawa, H. and J.C. Paulson (1994) J. Biol. Chem. 269:17872.
  5. Hennet, T. et al. (1998) Proc. Natl. Acad. Sci. USA 95:4504.
  6. Recchi, M.A. et al. (1998) Cancer Res. 58:4066.
  7. Dall’Olio, F. et al. (2001) Eur. J. Biochem. 268:5876.
  8. Weinstein, J. et al. (1987) J. Biol. Chem. 262:17735.
  9. Kitazume, S. et al. (2009) Glycobiology 19:479.
  10.  Wu, Z.L. et al. (2011) Glycobiology 21:727.

Long Name

ST6 beta-Galactoside alpha-2,6-Sialyltransferase 1

Alternate Names

Alpha 2,6-ST, B-cell antigen CD75, CD75, SIAT1, ST6N

Entrez Gene IDs

6480 (Human); 20440 (Mouse); 25197 (Rat)

Gene Symbol

ST6GAL1

UniProt

P15907

Additional ST6 Gal Sialyltransferase 1/ST6GAL1 Products

Product Documents for Recombinant Human ST6GAL1 (aa 44-406) 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.

Download SDS

Product Specific Notices for Recombinant Human ST6GAL1 (aa 44-406) Protein, CF

For research use only

Related Research Areas

Glycobiology-related Enzymes

Citations for Recombinant Human ST6GAL1 (aa 44-406) Protein, CF

Customer Reviews for Recombinant Human ST6GAL1 (aa 44-406) Protein, CF (1)

4 out of 5
1 Customer Rating
5 Stars
0%
4 Stars
100%
3 Stars
0%
2 Stars
0%
1 Stars
0%

Have you used Recombinant Human ST6GAL1 (aa 44-406) Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card
Showing  1 - 1 of 1 review Showing All
Filter By:
Clear Filters X
  • Recombinant Human ST6GAL1 (aa 44-406) Protein, CF
    Name: Sarah Groft
    Application: Enzymatic activity in vitro
    Verified Customer | Posted 03/22/2018

There are no reviews that match your criteria.

Showing  1 - 1 of 1 review Showing All

Protocols

View specific protocols for Recombinant Human ST6GAL1 (aa 44-406) Protein, CF (7620-GT):

Materials
  • Assay Buffer: 25 mM Tris, 150 mM NaCl, 5 mM CaCl2, 10 mM MnCl2, pH 7.5 (Tris and MnCl2 supplied in kit)
  • Recombinant Human Gal Sialyltransferase 1/ST6GAL1 (rhST6GAL1) (Catalog # 7620-GT)
  • CMP-Neu5Ac (Sigma, Catalog # C8271), 10 mM stock in deionized water
  • N-Acetyllactosamine (V-Labs, Catalog # GN204), 50 mM stock in deionized water
  • Sialyltransferase Activity Kit (Catalog # EA002)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • 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.
  2. 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 nmoles per well.
  3. Prepare reaction mixture containing 0.4 mM CMP-Neu5Ac, 2 mM N-Acetyllactosamine, and 4 µg/mL Coupling Phosphatase 2 in Assay Buffer.
  4. Dilute rhST6Gal1 to 20 μg/mL in Assay Buffer.
  5. Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of Assay Buffer.
  6. Load 25 µL of the 20 µg/mL rhST6GAL1 into the plate. Include a Control containing 25 µL of Assay Buffer.
  7. Add 25 µL of reaction mixture to the wells, excluding the standard curve.
  8. Cover the plate with parafilm or a plate sealer and incubate at 37 °C for 20 minutes.
  9. Add 30 µL of the Malachite Green Reagent A to all wells. Mix briefly.
  10. Add 100 µL of deionized water to all wells.
  11. Add 30 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
  12. Read plate at 620 nm (absorbance) in endpoint mode.
  13. 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:

  • rhST6GAL1: 0.5 µg
  • Coupling phosphatase 2: 0.1 µg
  • CMP-Neu5Ac: 0.2 mM
  • N-Acetyllactosamine: 1 mM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes