Recombinant Human SULT1E1 Protein, CF

R&D Systems | Catalog # 5545-ST

R&D Systems
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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human SULT1E1 Protein (5545-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

P49888

Applications

Enzyme Activity
View all Cytosolic Sulfotransferase 1E1/SULT1E1 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Cytosolic Sulfotransferase 1E1/SULT1E1 protein
Asn2-Ile294, with an N-terminal 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

His

Predicted Molecular Mass

36 kDa

SDS-PAGE

35 kDa, reducing conditions

Activity

Measured by its ability to transfer sulfate from PAPS to 1-Napthol.
The specific activity is >40 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

5545-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 1E1/SULT1E1

Cytosolic Sulfotransferases catalyze the sulfation 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. SULT1E1 is widely known as an estrogen sulfotransferase and may control estrogen levels by converting free estradiol to its inactive sulfate conjugate (1). Known substrates of this enzyme also include dehydroepiandrosterone, pregnenolone, ethinylestradiol, equalenin, diethylstilbesterol, 4-nitrophenol, 1-napthol and resveratrol (2, 3). SULT1E1 activity is found in reproductive organs (4), intestine and liver (3, 5). The enzymatic activity of the recombinant human SULT1E1 is measured using a phosphatasecoupled assay (6).

References

  1. Gong, H. et al. (2008) Cancer Res. 68:7386.
  2. Falany, C.N. et al. (1995) J. Steroid Biochem. Mol. Biol. 52:529.
  3. Miksits, M. et al. 2005, Xenobiotica 35:1101.
  4. Takase, Y. et al. (2007) The Prostate 67:405.
  5. Schrag, M.L. et al. (2004) Drug Metabolism and Disposition 32:1299.
  6. Prather, B. et al. (2012) Anal. Biochem. 423:86.

Alternate Names

EST-1, STE, SULT1E1

Entrez Gene IDs

6783 (Human); 20860 (Mouse); 25355 (Rat)

Gene Symbol

SULT1E1

UniProt

P49888

Additional Cytosolic Sulfotransferase 1E1/SULT1E1 Products

Product Documents for Recombinant Human SULT1E1 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 SULT1E1 Protein, CF

For research use only

Citations for Recombinant Human SULT1E1 Protein, CF

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Protocols

View specific protocols for Recombinant Human SULT1E1 Protein, CF (5545-ST):

Materials
  • Universal Sulfotransferase Activity Kit (Catalog # EA003)
  • 10X Assay Buffer (supplied in kit): 500 mM Tris, 150 mM MgCl2, pH 7.5
  • Recombinant human SULT-1E1 (rhSULT-1E1) (Catalog # 5545-ST)
  • 3'-Phosphoadenosine-5'-phosphosulfate (PAPS) (Catalog # ES019)
  • 1-Naphthol (Sigma, Catalog # N1000), 10 mM in deionized water
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Prepare 1X Assay Buffer by diluting 10X stock 10-fold with deionized water.
  2. 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 1X Assay Buffer for a 100 µM stock. This is the first point of the standard curve.
  3. Complete the standard curve by performing six one-half serial dilutions of the 100 µM Phosphate stock using 1X Assay Buffer. The standard curve has a range of 0.078 to 5 nmol per well.
  4. Prepare a reaction mixture containing 0.4 mM PAPS, 0.4 mM 1-Naphthol, and 0.02 mg/mL Coupling Phosphatase 3 in 1X Assay Buffer.
  5. Dilute rhSULT-1E1 to 20 µg/mL in 1X Assay Buffer.
  6. Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of 1X Assay Buffer.
  7. Load 25 µL of the 20 µg/mL rhSULT-1E1 into the plate. Include a Control containing 25µL of 1X 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 sealed plate 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 fitting and adjusted for Control.

Per Reaction:

  • rhSULT-1E1:  0.5 µg
  • PAPS: 10 nmoles (0.2 mM)
  • 1-Naphthol: 0.2 mM
  • Coupling Phosphatase 3: 0.5 µg

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