Recombinant Human SULT1C4 Protein, CF

R&D Systems | Catalog # 7095-ST

R&D Systems
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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human SULT1C4 Protein (7095-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

O75897

Applications

Enzyme Activity
View all Cytosolic Sulfotransferase 1C4/SULT1C4 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Cytosolic Sulfotransferase 1C4/SULT1C4 protein
Ala2-Phe302, with an N-terminal Met and 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

Met

Predicted Molecular Mass

36 kDa

SDS-PAGE

34-38 kDa, reducing conditions

Activity

Measured by its ability to transfer sulfate from PAPS to 1-Napthol.
The specific activity is >150 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

7095-ST
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, Brij-35 and DTT.
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: Cytosolic Sulfotransferase 1C4/SULT1C4

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). SULT1C4, originally designated as SULT1C2 (3), is expressed at higher levels in fetal lung and kidney and at lower levels in fetal heart. The endogenous substrate for this enzyme is unknown. It shows activity towards p‑nitrophenol and the carcinogenic compound, N‑hydroxy-2-acetylamino-fluorene (N-OH-2AAF) (3, 4).

References

  1. Falany, C. N. (1997) FASEB J. 11:206.
  2. Gamage, N. U. et al. (2006) Toxicol. Sci. 90:5.
  3. Sakakibara, Y. et al. (1998) J. Biol. Chem. 273:33929.
  4. Yoshinari, K. et al. (2001) J. Biochem. Mol. Toxicol. 15:67.

Alternate Names

ST1C4, SULT1C4

Entrez Gene IDs

27233 (Human)

Gene Symbol

SULT1C4

UniProt

O75897

Additional Cytosolic Sulfotransferase 1C4/SULT1C4 Products

Product Documents for Recombinant Human SULT1C4 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.

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Product Specific Notices for Recombinant Human SULT1C4 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 SULT1C4 Protein, CF (7095-ST):

Materials
  • Assay Buffer: 25 mM Tris, 15 mM MgCl2, pH 7.5 (Supplied in 10X form in kit)
  • Recombinant Human Cytosolic Sulfotransferase 1C4/SULT1C4 (rhSULT1C4) (Catalog # 7095-ST)
  • 3'-Phosphoadenosine-5'-phosphosulfate (PAPS) (Catalog # ES019)
  • 1-Naphthol (Sigma, Catalog # N1000), 10 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 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. 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. Dilute Coupling Phosphatase 3 to 0.125 mg/mL in Assay Buffer.
  4. Prepare reaction mixture containing 0.4 mM PAPS, 1.12 mM 1-Naphthol, and 20 μg/mL Coupling Phosphatase 3.
  5. Dilute rhSULT1C4 to 6 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 6 µg/mL rhSULT1C4 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. Cover the plate with a plate sealer 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.

Per Reaction:

  • rhSULT1C4: 0.15 µg
  • PAPS: 0.2 mM
  • 1-Naphthol: 0.56 mM
  • Coupling Phosphatase 3: 0.5 µg

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