Recombinant Human SULT2A1 Protein, CF

R&D Systems | Catalog # 5828-ST

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human SULT2A1 Protein (5828-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

Q06520

Applications

Enzyme Activity
View all Cytosolic Sulfotransferase 2A1/SULT2A1 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Cytosolic Sulfotransferase 2A1/SULT2A1 protein
Ser2-Glu285, with an N-terminal Met and 6-His tag

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.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

35 kDa

SDS-PAGE

36 kDa, reducing conditions

Activity

Measured by its ability to transfer sulfate from PAPS to t-Dehydroandrosterone (DHEA).
The specific activity is >15 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

5828-ST
Formulation Supplied as a 0.2 μm filtered solution in Tris, 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 2A1/SULT2A1

Cytosolic sulfotransferases are a family of Phase II drug-metabolizing enzymes that catalyze the sulfation of many endogenous and xenobiotic substrates (1‑3). They have important roles in the metabolism of many endogenous compounds including steroids, bile acids, thyroid hormones and monoamine neurotransmitters. They are distributed throughout the body and serve to inactivate and increase water-solubility of xenobiotics and therapeutic drugs. They are distinct from Golgi resident sulfotransferases by lacking N-terminal signal-anchor domains and residing only in the cytoplasm. SULT2A1 is primarily expressed in liver and adrenal tissues where it sulfates steroids and bile acids (4‑6). It is also called DHEA/bile acid sulfotransferase and plays roles in maintaining cholesterol and bile acid homeostasis by increasing cholesterol catabolism and, at the same time, preventing toxicity from bile acid accumulation. The enzymatic activity of the recombinant human SULT2A1 is measured using a phosphatase-coupled assay (7).

References

  1. Falany, C. N. (1997) FASEB J. 11:206.
  2. Gamage, N. U. et al. (2006) Toxicol. Sci. 90:5.
  3. Allali-Hassani, A. et al. (2007) PLoS Biol. 5:e97.
  4. Comer, K. A. et al. (1993) Biochem. J. 289:233.
  5. Otterness, D.M. et al. (1992) Mol. Pharmacol. 41:865.
  6. Luu-The, V. et al. (1995) DNA Cell Biol. 14: 511.
  7. Prather, B. et al. (2012) Anal. Biochem. 423:86.

Alternate Names

DHEA-ST, DHEAS, hSTa, ST2A3, SULT2A1

Entrez Gene IDs

6822 (Human); 20859 (Mouse); 24912 (Rat)

Gene Symbol

SULT2A1

UniProt

Q06520

Additional Cytosolic Sulfotransferase 2A1/SULT2A1 Products

Product Documents for Recombinant Human SULT2A1 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human SULT2A1 Protein, CF

For research use only

Citations for Recombinant Human SULT2A1 Protein, CF

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Protocols

View specific protocols for Recombinant Human SULT2A1 Protein, CF (5828-ST):

Materials
  • Assay Buffer: 50 mM Tris, 15 mM MgCl2, pH 7.5 (supplied in 10X form in kit)
  • Recombinant Human Cytosolic Sulfotransferase 2A1/SULT2A1 (rhSULT2A1) (Catalog # 5828-ST)
  • 3'-Phosphoadenosine-5'-phosphosulfate (PAPS) (Catalog # ES019)
  • T-Dehydroandrosterone (DHEA)  (Sigma, Catalog # D4000), 1 mM in 50% DMSO, 50% deionized water
  • Universal Sulfotransferase Activity Kit (Catalog # EA003)
  • 96-well Clear Plate (Catalog # DY990)
  • 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 140 µL of 1 mM PAPS, 70 µL of 1 mM DHEA, 70 µL of 0.1 mg/mL Coupling Phosphatase 3, and 70 µL of Assay Buffer.
  3. Dilute rhSULT2A1 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 rhSULT2A1 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 parafilm or 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:

  • rhSULT2A1: 1.0 µg
  • PAPS: 10000 pmoles (0.2 mM)
  • DHEA: 0.1 mM
  • Coupling Phosphatase 3: 0.5 µg

FAQs

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