>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
<1.0 EU per 1 μg of the protein by the LAL method.
39 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
55-75 kDa, reducing conditions
Read Publication using 6140-ST in the following applications:
Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
Dilute 1 mM Phosphate Standard provided by the Universal Sulfotransferase Kit by adding 40 µL of the 1 mM Phosphate Standard to 360 µL of Assay Buffer for a 100 µM stock.
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 nmol per well.
Prepare reaction mixture containing 0.4 mM PAPS, 1 mM PGA, and 20 μg/mL Coupling Phosphatase 3 in Assay Buffer.
Dilute rhCHST10 to 4 µg/mL in Assay Buffer.
Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of Assay Buffer.
Load 25 µL of the 4 µg/mL rhCHST10 into the plate. Include a Control containing 25 µL of Assay Buffer.
Add 25 µL of reaction mixture to the wells, excluding the standard curve.
Cover the plate with a plate sealer and incubate at 37 °C for 20 minutes.
Add 30 µL of the Malachite Green Reagent A to all wells. Mix briefly.
Add 100 µL of deionized water to all wells. Mix briefly.
Add 30 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
Read plate at 620 nm (absorbance) in endpoint mode.
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.
rhCHST10: 0.1 μg
Coupling Phosphatase 3: 0.5 μg
PAPS: 0.2 mM
PGA: 0.5 mM
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human CHST10 Protein, CF
Carbohydrate Sulfotransferase 10
CHST10 is the only known sulfotransferase that catalyzes the transfer of sulfate to position 3 of terminal glucuronic acid of both protein- and lipid-linked oligosaccharides (1, 2). CHST10 is a key enzymes for the synthesis of HNK-1 carbohydrate structure, a sulfated glucuronyl-lactosaminyl residue carried by many neural recognition molecules (3). HNK-1 is involved in cell interactions during ontogenetic development and in synaptic plasticity in the adult. In adults, CHST10 is highly expressed in brain, testis and ovary. Recently, it is reported that CHST10 functions as a suppressor of invasiveness of human melanoma cells (4). Like many other carbohydrate specific sulfotransferases, CHST10 is a single-pass type II membrane protein and is localized in Golgi apparatus membrane. The recombinant human CHST10 only contains the luminal enzymatic domain and the enzymatic activity is measured using a phosphatase-coupled assay (5).
Ong, E. et al. (1999) J. Biol. Chem. 274:25608.
Ong, E. et al. (1998) J. Biol. Chem. 273:5190.
Kizuka, Y. et al. (2006) J. Biol. Chem. 281:13644.
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