Recombinant Human Heparan Sulfate-6-O-Sulfotransferase-1, CF

  (2 citations)     
Datasheet / CoA / SDS
Product Details
Citations (2)
FAQs
Supplemental Products
Reviews
  • Purity
    >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
  • Endotoxin Level
    <1.0 EU per 1 μg of the protein by the LAL method.
  • Activity
    Measured by its ability to transfer sulfate from PAPS to heparan sulfate. The specific activity is >45 pmol/min/μg, as measured under the described conditions.
  • Source
    Chinese Hamster Ovary cell line, CHO-derived human Heparan Sulfate 6-O-Sulfotransferase 1/HS6ST1 protein
    Pro28-Trp401, with an N-terminal 6-His tag
  • Accession #
  • N-terminal Sequence
    Analysis
    His
  • Predicted Molecular Mass
    45 kDa
  • SDS-PAGE
    56-60 kDa, reducing conditions
Product Datasheets

Certificate of Analysis Lookup

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Certificate of Analysis Lookup

Certificate of Analysis Request Form

To download a Certificate of Analysis, please enter the catalog and lot numbers in the search box below.

Note: Certificate of Analysis not available for kit components.

5057-ST
 
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and EDTA.
 
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.
 
Assay Procedure
Materials
  • Assay Buffer: 50 mM Tris, 15 mM MgCl2, pH 7.5
  • Recombinant Human Heparan Sulfate 6‑O‑Sulfotransferase 1/HS6ST1 (rhHS6ST1) (Catalog # 5057-ST)
  • Donor Substrate: 3'-Phosphoadenosine-5'-phosphosulfate (PAPS) (Catalog # ES019)
  • Acceptor Substrate: Heparan Sulfate (Celsus Labs, Catalog # HO3105), 50 mg/mL stock in deionized water
  • Sulfotransferase Activity Kit (Catalog # EA003)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Create Assay Buffer by Diluting 10X Phosphatase Buffer 3 (supplied in kit) to 1X in deionized water.
  2. Dilute 1 mM Phosphate Standard by adding 40 µL of the 1 mM Phosphate Standard to 360 µL of Assay Buffer for a 100 µM stock.
  3. Continue standard curve by performing seven one-half serial dilutions of the 100 µM Phosphate stock in Assay Buffer. The standard curve has a range of 0.039 to 2.5 nmol per well.
  4. Prepare a reaction mixture containing 0.4 mM PAPS, 4 mg/mL Heparin Sulfate, and 20 ug/mL Coupling Phosphatase 3 in Assay Buffer. *Note: Incorporate Coupling Phoshphatase 3 into reaction mixture right before the mixture is added to the plate in order to avoid unnecessary hydrolysis of PAPS.
  5. Dilute rhHS6ST1 to 20 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 Assay Buffer.
  7. Load 25 µL of the 40 µg/mL rhHS6ST1 into the plate. Include a control containing 25 µL of 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.
  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) =

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:
  • rhHS6ST1: 0.5 µg
  • Coupling Phosphatase 3: 0.5 µg
  • Heparan Sulfate: 100 ug (2 mg/mL)
  • PAPS: 0.2 mM
Background: Heparan Sulfate 6-O-Sulfotransferase 1/HS6ST1

Heparan Sulfate is a highly sulfated polysaccharide that can be found on cell surface and extracellular matrix. It is usually covalently attached to a protein core as the glycan component of a proteoglycan. Heparan Sulfate interacts with a variety of proteins, such as growth factors, protease inhibitors, cytokines, lipoprotein lipase and viral envelope proteins, thus plays roles from cell growth, cell differentiation, cell motility, blood coagulation, lipid metabolism to viral infection (1, 2). Heparan Sulfate consists of repeating residues of uronic acid and N‑acetylglucosamine. The uronic acid residues can be sulfated at 2-O position by Heparan Sulfate 2‑O‑Sulfotransferase (HS2ST). The N‑acetylglucosamine residues can be sulfated at N-, 3-O, and 6-O positions by N‑deacetylase/N‑sulfotransferases (NDSTs), Heparan Sulfate 3-O and 6-O sulfotransferases (HS3STs and HS6STs) respectively. However, the reactions catalyzed by these sulfotransferases are normally incomplete on the whole chain of Heparan Sulfate. As a result, Heparan Sulfate displays enormous sequence diversity that allows it to interact with a wide spectrum of proteins differently. Among three HS6STs, HS6ST1 is the first to be cloned (3). Mice deficient of the HS6ST1 homologue gene showed embryonic lethality (4).The enzymatic activity of the recombinant human HS6ST1 is measured using a phosphatase-coupled assay (5).

  • References:
    1. Bernfield, M. et al. (1999) Annu. Rev. Biochem. 68:729.
    2. Esko, J. D. and Selleck, S. B. (2002) Annu. Rev. Biochem. 71:435.
    3. Habuchi, H et al. (1998) J. Biol. Chem. 273:9208.
    4. Habuchi, H et al. (2007) J. Biol. Chem. 282:15578.
    5. Prather, B. et al. (2012) Anal. Biochem. 423:86.
  • Entrez Gene IDs:
    9394 (Human); 50785 (Mouse); 316325 (Rat)
  • Alternate Names:
    DKFZp547H098; EC 2.8.2; EC 2.8.2.-; FLJ25392; heparan sulfate 6-O-sulfotransferase 1; heparan-sulfate 6-sulfotransferase; HS6ST1; HS6ST-1; HS6STheparan-sulfate 6-O-sulfotransferase 1; MGC116899; MGC116901
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
Filter your results:

Species
Applications
Sample Type
  1. Structural and functional study of D-glucuronyl C5-epimerase.
    Authors: Qin Y, Ke J, Gu X, Fang J, Wang W, Cong Q, Li J, Tan J, Brunzelle J, Zhang C, Jiang Y, Melcher K, Li J, Xu H, Ding K
    J Biol Chem, 2015;290(8):4620-30.
    Species: Zebrafish
    Sample Type: Protein
    Application: Bioassay
  2. Detection of specific glycosaminoglycans and glycan epitopes by in vitro sulfation using recombinant sulfotransferases.
    Authors: Wu ZL, Prather B, Ethen CM
    Glycobiology, 2011;21(5):625-33.
    Species: Bovine
    Sample Type: Protein
    Application: EnzAct

FAQs

No product specific FAQs exist for this product, however you may

View all Proteins and Enzyme FAQs
Primary Antibodies
Description Application Cat# Citations Images  

His Tag Antibody

WB , Simple Western , Flow , AP , CyTOF-ready MAB050 32  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

His Tag HRP-conjugated Antibody

WB , Simple Western MAB050H 3  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

His Tag PE-conjugated Antibody

ICFlow IC050P 4  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

His Tag APC-conjugated Antibody

ICFlow IC050A  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

His Tag Antibody

WB , Flow , AP MAB050R 2  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

His Tag Biotinylated Antibody

WB BAM050 4  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

His Tag Alexa Fluor® 488-conjugated Antibody

ICFlow IC050G  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

His Tag Alexa Fluor® 700-conjugated Antibody

ICFlow IC050N  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

His Tag PerCP-conjugated Antibody

ICFlow IC050C  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....
Protein Purification Kits
Description Application Cat# Citations Images  

Histidine-tagged Protein Purification Resin

IP999 1
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....
Sulfotransferase Assays and Substrates
Description Application Cat# Citations Images  

3'-Phosphoadenosine-5'-phosphosulfate

ES019 2
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Have you used Recombinant Human Heparan Sulfate-6-O-Sulfotransferase-1, CF?

Submit a review and receive a $25US/€18/£15/$25CAN amazon gift card if you include an image - $10US/€7/£6/$10CAN Amazon card for reviews without an image. Limited to verified customers in USA, Canada and Europe.

Order Details

Contact Us to Order
reload

Feedback

Customer Information
Calculators

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

=
÷
R&D Systems Guarantee