Recombinant Human SULT1A3 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human SULT1A3 Protein, CF Summary

Details of Functionality
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.
Source
E. coli-derived human Cytosolic Sulfotransferase 1A3/SULT1A3 protein
Glu2-Leu295 with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Met
Protein/Peptide Type
Recombinant Enzymes
Gene
SULT1A3
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
35 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.
SDS-PAGE
36 kDa, reducing conditions
Publications
Read Publication using
5829-ST in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol and DTT.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 50 mM Tris, 15 mM MgCl2, pH 7.5
  • Recombinant Human Sulfotransferase 1A3/SULT1A3 (rhSULT1A3) (Catalog # 5829-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.
  2. Prepare a 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.
  3. Prepare a reaction mixture containing 0.4 mM PAPS, 0.4 mM 1-Naphthol, and 0.02 mg/mL Coupling Phosphatase 3 in Assay Buffer.
  4. Dilute rhSULT1A3 to 10 µg/mL in Assay Buffer.
  5. Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of Assay Buffer.
  6. Load 25 µL of the diluted rhSULT1A3 into the plate. Include a Substrate Blank containing 25 µL of Assay Buffer.
  7. Add 25 µL of reaction mixture to the wells, excluding the standard curve.
  8. Cover the plate with a plate sealer and incubate at 37 °C for 20 minutes.
  9. Add 30 µL of the Malachite Green Reagent A to all wells. Mix briefly.
  10. Add 100 µL of deionized water to all wells. Mix briefly.
  11. Add 30 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
  12. Read plate at 620 nm (absorbance) in endpoint mode.
  13. 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 Substrate Blank.

Per Reaction:
  • rhSULT1A3: 0.25 µg
  • PAPS: 10,000 pmol (0.2 mM)
  • 1-Naphthol: 0.2 mM
  • Coupling Phosphatase 3: 0.5 µg

Notes

Glogos is registered trademark of Stratagene.



This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human SULT1A3 Protein, CF

  • Cytosolic Sulfotransferase 1A3
  • EC 2.8.2
  • EC 2.8.2.1
  • HAST
  • HAST3
  • M form) phenol sulfotransferase
  • MGC117469
  • Monoamine-sulfating phenol sulfotransferase
  • M-PST
  • ST1A3/ST1A4
  • STM
  • sulfotransferase family, cytosolic, 1A, phenol-preferring, member 3
  • Sulfotransferase, monoamine-preferring
  • SULT1A3
  • Thermolabile phenol sulfotransferase
  • TL-PST

Background

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 widely 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.  SULT1A3 catalyzes the sulfation of phenolic monoamines, such as dopamine, norepinephrine and serotonin, and phenolic and catecholic drugs (4, 5). SULT1A3 is also called catecholamine sulfotransferase. The enzymatic activity of the recombinant human SULT1A3 is measured using a phosphatasecoupled assay (6).

  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. Dooley, T.P. et al. (1994) Biochem. Biophys. Res. Commun. 205:1325.
  5. Salman, E.D. et al. (2009) Drug Metab. Dispos. 37:706.
  6. Prather, B. et al. (2012) Anal. Biochem. 423:86. 
 

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5829-ST
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Publications for Cytosolic Sulfotransferase 1A3/SULT1A3/CSPS (5829-ST)(1)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 1 application: EnzAct.


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Bioinformatics

Gene Symbol SULT1A3
Entrez
Uniprot