Recombinant Human SULT1A1 Protein, CF Summary
Glu2-Leu295, with an N-terminal Met and 6-His tag
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|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.
- Assay Buffer: 50 mM Tris, 15 mM MgCl2, pH 7.5 (1X Phosphatase Buffer 3)
- Recombinant Human Cytosolic Sulfotransferase 1A1/SULT1A1 (rhSULT1A1) (Catalog # 5546-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
- Dilute 1 mM Phosphate Standard by combining 40 µL with 360 µL of Assay Buffer for a 100 µM stock. This is the first standard curve point.
- Prepare additional points 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.
- 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.
- Dilute rhSULT1A1 to 20 µg/mL in Assay Buffer.
- Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of Assay Buffer.
- Load 25 µL of the 20 µg/mL rhSULT1A1 into the plate. Include a Control containing 25 µL of Assay Buffer.
- Add 25 µL of reaction mixture to the wells, excluding the standard curve.
- Cover the plate with a plate sealer and incubate at 37 °C for 20 minutes.
- Add 30 µL of the Malachite Green Reagent A to all wells. Mix briefly.
- Add 100 µL of deionized water to all wells. Mix briefly.
- Add 30 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
- Read plate at 620 nm (absorbance) in endpoint mode.
- 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:
- rhSULT1A1: 0.5 µg
- PAPS: 10,000 pmol (0.2 mM)
- 1-Naphthol: 0.2 mM
- Coupling Phosphatase 3: 0.5 µg
Background: Cytosolic Sulfotransferase 1A1/SULT1A1
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). SULT1A1 is one of two phenol sulfotransferases with thermostable enzyme activity (2). The enzyme is more specific for sulfonation on catecholamines, phenolic drugs and neurotransmitters (3). Because of its ability to modify diverse promutagen and procarcinogen xenobiotics, it is implicated in a range of cancers (2). The crystal structure of the enzyme reveals that its active site is flexible and can accommodate diverse hydrophobic substrates with varying sizes, shapes and flexibility (4). The enzymatic activity of the recombinant human SULT1A1 is measured using a phosphatase-coupled assay (5).
- Falany, C. N. (1997) FASEB J. 11:206.
- Gamage, N. U. et al. (2006) Toxicol. Sci. 90:5.
- Wilborn, T.W. et al. (1993) Mol. Pharm. 43:70.
- Gamage, N. U. et al. (2003) J. Biol. Chem. 278:7655.
- Prather, B. et al. (2012) Anal. Biochem. 423:86.
Citations for Recombinant Human SULT1A1 Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
Citations: Showing 1 - 3
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LC-MS/MS quantification of sulfotransferases is better than conventional immunogenic methods in determining human liver SULT activities: implication in precision medicine
Authors: C Xie, TM Yan, JM Chen, XY Li, J Zou, LJ Zhu, LL Lu, Y Wang, FY Zhou, ZQ Liu, M Hu
Sci Rep, 2017;7(1):3858.
Sample Types: Protein
Applications: Enzyme Assay
Correlating chemical sensitivity and basal gene expression reveals mechanism of action.
Authors: Rees M, Seashore-Ludlow B, Cheah J, Adams D, Price E, Gill S, Javaid S, Coletti M, Jones V, Bodycombe N, Soule C, Alexander B, Li A, Montgomery P, Kotz J, Hon C, Munoz B, Liefeld T, Dancik V, Haber D, Clish C, Bittker J, Palmer M, Wagner B, Clemons P, Shamji A, Schreiber S
Nat Chem Biol, 2016;12(2):109-16.
Sample Types: Recombinant Protein
A versatile polyacrylamide gel electrophoresis based sulfotransferase assay.
Authors: Wu ZL, Ethen CM, Larson S, Prather B, Jiang W
BMC Biotechnol., 2010;10(0):11.
Sample Types: Recombinant Protein
Applications: Enzyme Assay
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Sulfotransferase Assays and Substrates
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