Recombinant Human SULT1C2 Protein, CF

R&D Systems | Catalog # 7458-ST

R&D Systems
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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human SULT1C2 Protein (7458-ST)
  • 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

E. coli

Accession Number

O00338

Applications

Enzyme Activity
View all Cytosolic Sulfotransferase 1C2/SULT1C2 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Cytosolic Sulfotransferase 1C2/SULT1C2 protein
Ala2-Leu296, with N-terminal 6-His tag

Purity

>80%, 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

Inconclusive. Protein identity confirmed by mass spec, His tag confirmed by anti-His Western analysis.

Predicted Molecular Mass

38 kDa

SDS-PAGE

34-37 kDa, reducing conditions

Activity

Measured by its ability to transfer sulfate from PAPS to 4-nitrophenol.
The specific activity is >45 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

7458-ST
Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl and DTT.
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.

Background: Cytosolic Sulfotransferase 1C2/SULT1C2

Cytosolic sulfotransferases catalyze the sulfonation of many hormones, neurotransmitters, drugs, and xenobiotic compounds. They are distinct from Golgi-resident sulfotransferases by the absence of transmembrane domains and are located in the cytoplasm (1, 2). SULT1C2 is mainly expressed in the gastrointestinal tract (stomach, duodenum, jejunum, ileum, colon, caecum and rectum), liver and kidneys, but not in the lungs (3). In contrast, SULT1C4, a sulfotransferase that is most closely related to SULT1C2 (4), is expressed at higher levels in fetal lung and kidney and at lower levels in fetal heart. So far, SULT1C2 is found to be active only on p-nitrophenol (3). The enzymatic activity of our recombinant human SULT1C2 was determined using a phosphatase-coupled assay (5).

References

  1. Falany, C. N. (1997) FASEB J. 11:206.
  2. Gamage, N. U. et al. (2006) Toxicol. Sci. 90:5.
  3. Hehonah, N. et al. (1999) Int J. Biochem. Cell. Biol. 31:869.
  4. Sakakibara, Y. et al. (1998) J. Biol. Chem. 273:33929.
  5. Prather, B. et al. (2012) Anal. Biochem. 423:86.

Alternate Names

ST1C1, ST1C2, SULT1C1, SULT1C2

Entrez Gene IDs

6819 (Human); 316153 (Rat)

Gene Symbol

SULT1C2

UniProt

O00338 (Human)

Additional Cytosolic Sulfotransferase 1C2/SULT1C2 Products

Product Documents for Recombinant Human SULT1C2 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human SULT1C2 Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

Related Research Areas

Sulfation Regulators

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Protocols

View specific protocols for Recombinant Human SULT1C2 Protein, CF (7458-ST):

Materials
  • Assay Buffer (1X Phosphatase Buffer 3): 50 mM Tris, 15 mM MgCl2, pH 7.5
  • Recombinant Human Cytosolic Sulfotransferase 1C2/SULT1C2 (rhSULT1C2) (Catalog # 7458-ST)
  • Coupling Enzyme: Recombinant Mouse IMPAD1 (Catalog # 7028-PD)
  • Adenosine 3′-phosphate 5′-phosphosulfate lithium salt hydrate (PAPS) (Catalog # ES019), 1 mM stock in 5% ethanol, 95% deionized water
  • 4-Nitrophenol (Sigma, Catalog # 241326), 50 mM in deionized water
  • Universal Sulfotransferase Activity Kit (Catalog # EA003)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute Coupling Phosphatase 3 to 0.1 mg/mL in Assay Buffer.
  2. Prepare reaction mixture by combining 50 µL of 0.1 mg/mL Coupling Phosphatase 3, 50 µL of 50 mM 4-Nitrophenol, 50 µL of 1 mM PAPS, and 100 µL of Assay Buffer. This is sufficient to assay 9 wells.
  3. Dilute rhSULT1C2 to 40 µg/mL in Assay Buffer.
  4. 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.
  5. 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.
  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 rhSULT1C2 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. Mix briefly.
  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.

  • rhSULT1C2:  1.0 µg
  • Coupling Phosphatase 3:  0.5 µg
  • 4-Nitrophenol:  5 mM
  • PAPS:  0.1 mM

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