Recombinant Human Exostosin 1 Protein, CF

Catalog #: 8379-GT Datasheet / COA / SDS
Newer Version Available: 8567-GT

Discontinued Product

8379-GT has been discontinued and is replaced by 8567-GT.

R&D Systems Recombinant Proteins and Enzymes
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Recombinant Human Exostosin 1 Protein, CF Summary

Product Specifications

>90%, 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.
Measured by its ability to transfer GlcNAc and GlcA from donor substrates to heparan sulfate. The specific activity is >300 pmol/min/μg, as measured under the described conditions.
Chinese Hamster Ovary cell line, CHO-derived human Exostosin 1 protein
Asp69-Leu746 with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Asp69 & Leu71
Predicted Molecular Mass
79 kDa
65-84 kDa, reducing conditions

Product Datasheets

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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
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.

Assay Procedure

  • Assay Buffer: 25 mM Tris, 150 mM NaCl, 5 mM CaCl2,  5 mM MnCl2, pH 7.5
  • Recombinant human EXT1 (rhEXT1) (Catalog # 8379-GT)
  • UDP-GlcA (Sigma, Catalog # U5625), 10 mM stock in deionized water
  • UDP-GlcNAc (Sigma, Catalog # U4375), 50 mM stock in 50% ethanol/50% deionized water
  • Heparan Sulfate (Celsus Labs, Catalog # HO-03102 or HO-3105), 50 mg/mL stock in deionized water
  • Glycosyltransferase Activity Kit (Catalog # EA001)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. 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. This is the first point of the standard curve.
  2. 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.
  3. Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of Assay Buffer.
  4. Prepare a Reaction Mixture composed of 1 mM UDP-GlcA, 1 mM UDP-GlcNAc, 8 mg/mL Heparan Sulfate, and 4 µg/mL Coupling Phosphatase 1 in Assay Buffer.
  5. Dilute rhEXT1 to 4 µg/mL in Assay Buffer.
  6. Load 25 µL of the 4 µg/mL rhEXT1 into the plate. Include a Substrate Blank containing 25 µL of Assay Buffer.
  7. Start the reaction by adding 25 µL of Reaction Mixture to the wells, excluding the standard curve and curve blank.
  8. Cover the plate with 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. Mix briefly.
  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:
  • rhEXT1: 0.1 µg
  • UDP-GlcA: 0.5 mM
  • UDP-GlcNAc: 0.5 mM
  • Heparan Sulfate: 200 µg
  • Coupling Phosphatase 1: 0.1 µg
  • Reconstitution Calculator

    Reconstitution Calculator

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    Background: Exostosin 1

    The exostosin (EXT) family of glycosyltransferases are involved in heparan sulfate (HS) and heparin biosynthesis (1). Mutations of these enzymes are the causes of hereditary multiple exostoses, which are characterized by formation of numerous cartilage-capped, benign bone tumors (osteocartilaginous exostoses or osteochondromas) that are often accompanied by skeletal deformities and short stature (2). In a small percentage of cases exostoses have exhibited malignant transformation resulting in an osteosarcoma or chondrosarcoma (3). Five members of this family have been cloned to date: EXT1, EXT2, EXTL1, EXTL2, and EXTL3. EXT1 and EXT2 are bifunctional enzymes with both beta -1,4-glucuronyltransferase and alpha -1,4-N-acetylglucosaminyltransferase activities that add alternating beta -1,4-GlcA and alpha -1,4-GlcNAc residues on the non-reducing end of HS chain (4). EXT1 is an active enzyme, but the hetero-dimer of EXT1 and EXT2 is more stable and has higher activity (1). The enzyme activity of recombinant human EXT1 is measured using a phosphatase-coupled assay (5).

    1. Busse, M. et al. (2007) J. Biol. Chem. 282:32802.
    2. Wuyts, W. et al. (1998) Am. J. Hum. Genet. 62:346.
    3. Hecht J.T. et al. (1997) Am. J. Hum. Genet. 60:80.
    4. Kobayashi S. et al. (2000) Biochem. Biophys. Res. Commun. 268:860.
    5. Wu, Z.L. et al. (2011) Glycobiology 21:727.
    Long Name
    Exostosin Glycosyltransferase 1
    Entrez Gene IDs
    2131 (Human); 14042 (Mouse); 299907 (Rat)
    Alternate Names
    EC; EC; exostoses (multiple) 1; Exostosin 1; exostosin-1; EXT; EXT1; Glucuronosyl-N-acetylglucosaminyl-proteoglycan 4-alpha-N-acetylglucosaminyltransferase; Glucuronosyl-N-acetylglucosaminyl-proteoglycan/N-acetylglucosaminyl-proteoglycan4-alpha-N-acetylglucosaminyltransferase; Langer-Giedion syndrome chromosome region; LGCR; LGS; Multiple exostoses protein 1; N-acetylglucosaminyl-proteoglycan 4-beta-glucuronosyltransferase; Putative tumor suppressor protein EXT1; TRPS2; TTV


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