Recombinant Human B3GNT6 Protein, CF

R&D Systems | Catalog # 6505-GT

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems CHO-derived Recombinant Human B3GNT6 Protein (6505-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

Q6ZMB0

Applications

Bioactivity
View all Beta-1,3-N-Acetylglucosaminyltransferase 6/B3GNT6 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Beta-1,3-N-Acetylglucosaminyltransferase 6/B3GNT6 protein
Gln32-Ser384, 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

40 kDa

SDS-PAGE

43-55 kDa, reducing conditions

Activity

Measured by its ability to transfer GlcNAc from UDP-GlcNAc to 4-nitrophenyl-alpha -D-galactosaminide.
The specific activity is >35 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6505-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: Beta-1,3-N-Acetylglucosaminyltransferase 6/B3GNT6

beta -1,3-N-acetylglucosaminyltransferase 6 (B3GNT6) is also known as core 3 synthase due to its role in synthesis of the core 3 structure (GlcNAc beta 1‑3Gal‑NAc alpha 1‑serine/threonine), an important precursor in the biosynthesis of mucin-type glycoproteins in digestive organs (1). Its expression is restricted to the stomach, colon and small intestine, where the core 3 structure is present. Down-regulation of the enzyme was found in gastric and colorectal carcinomas and it was suggested that it may be useful as a marker to distinguish between benign adenomas and premalignant lesions (2, 3). Prostate cancer cells transfected with core 3 synthase exhibited reduced migration and invasion compared with mock-transfected cells (4). When inoculated into nude mice, the transfected cells produced smaller tumors without metastasis in contrast to the robust tumor formation and metastasis observed in mock-transfected cells (4). Like other members of the beta ‑1,3‑N‑acetylglucosaminyltransferase family, B3GNT6 is a Golgi-resident single-pass type II membrane protein.The activity of this enzyme has been measured with a phosphatase-coupled method (5).

References

  1. Iwai, T. et al. (2002) J. Biol. Chem. 277:12802.
  2. Iwai, T. et al. (2005) Proc. Natl. Acad. Sci. USA 102:4572.
  3. Vavasseur, F. et al. (1995) Glycobiology 5:351.
  4. Lee, S.H. et al. (2009) J. Biol. Chem. 284:17157.
  5. Wu, Z.L. et al. (2010) Glycobiology doi: 10.1093/glycob/cwq187.

Long Name

UDP-GlcNAc:betaGal beta-1,3-N-acetylglucosaminyltransferase 6

Alternate Names

Beta3Gn-T6, BGnT-6, Core 3 synthase

Entrez Gene IDs

192134 (Human)

Gene Symbol

B3GNT6

UniProt

Q6ZMB0

Additional Beta-1,3-N-Acetylglucosaminyltransferase 6/B3GNT6 Products

Product Documents for Recombinant Human B3GNT6 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 B3GNT6 Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

Related Research Areas

Glycobiology-related Enzymes

Citations for Recombinant Human B3GNT6 Protein, CF

Customer Reviews for Recombinant Human B3GNT6 Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human B3GNT6 Protein, CF and earn rewards!

Have you used Recombinant Human B3GNT6 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

Protocols

View specific protocols for Recombinant Human B3GNT6 Protein, CF (6505-GT):

Materials
  • Assay Buffer: 25 mM Tris, 150 mM NaCl, 10 mM MnCl2, 5 mM CaCl2, 20% DMSO, pH 7.5
  • Recombinant Human beta -1,3-N-acetylglucosaminyltransferase 6/B3GNT6 (rhB3GNT6) (Catalog # 6505-GT)
  • Coupling Enzyme: Recombinant Human CD39L3/ENTPD3 (rhCD39L3) (Catalog # 4400-EN)
  • UDP-GlcNAc (Sigma, Catalog # U4375), 50 mM stock in 50% EtOH (v/v)
  • 4-Nitrophenyl N-acetyl-alpha -D-galactosaminide (4-NP-GalNAc) (Sigma, Catalog # N4264), 15 mM stock in DMSO
  • Malachite Green Phosphate Detection Kit (Catalog # DY996)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute UDP-GlcNAc to 1.2 mM in Assay Buffer.
  2. Dilute 4-NP-GalNAc to 3.6 mM in Assay Buffer.
  3. Dilute rhCD39L3 to 6 μg/mL in Assay Buffer.
  4. Prepare reaction mixture by combining equal volumes of 1.2 mM UDP-GlcNAc, 3.6 mM 4-NP-GalNAc, and 6 μg/mL rhCD39L3.
  5. Dilute rhB3GNT6 to 12 µg/mL in Assay Buffer.
  6. Dilute 1 M Phosphate Standard by adding 10 µL of the 1 M Phosphate Standard to 990 µL of deionized water for a 10 mM stock. Continue by adding 10 µL of the 10 mM Phosphate stock to 990 µL of Assay Buffer for a 100 µM stock. This is the first point of the standard curve.
  7. Continue 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 5.0 nmol per well.
  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 12 µg/mL rhB3GNT6 into the plate. Include a substrate blank 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 ºC for 60 minutes.
  12. Add 30 µL of the Malachite Green Reagent A to all wells. Mix and incubate for 10 minutes at room temperature.
  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 Substrate.

Per Reaction:

  • rhB3GNT6: 0.300 µg
  • rhCD39L3: 50 ng
  • UDP-GlcNAc: 0.2 mM
  • 4-NP-GalNAc: 0.6 mM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes