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Recombinant Human Aldo-keto Reductase 1C4/AKR1C4 Protein, CF

Catalog #: 6957-DH Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
6957-DH-020
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Recombinant Human Aldo-keto Reductase 1C4/AKR1C4 Protein, CF Summary

Product Specifications

Purity
>90%, 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.
Activity
Measured by its NADP+-dependent oxidation of cholic acid. The specific activity is >300 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived human Aldo-keto Reductase 1C4/AKR1C4 protein
Met1-Tyr323
Accession #
NP_001809
N-terminal Sequence
Analysis
Met1
Predicted Molecular Mass
37 kDa
SDS-PAGE
36-38 kDa, reducing conditions

Product Datasheets

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6957-DH

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

6957-DH

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol 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.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Tris, 500 mM NaCl, pH 8.5
  • Recombinant Human Aldo-keto Reductase 1C4/AKR1C4 (rhAKR1C4) (Catalog # 6957-DH)
  • Sodium Cholate (Sigma, Catalog # C1254), 464.5 mM (20% w/v) stock in deionized water
  • beta -Nicotinamide adenine dinucleotide phosphate (NADP+) (Sigma, Catalog # N5755), 50 mM stock in deionized water
  • UV Plate (Costar, Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhAKR1C4 to 6 ng/μL in Assay Buffer.
  2. Prepare a Reaction Mixture containing 2 mM sodium cholate and 1 mMNADP+ in Assay Buffer.
  3. In a plate, load 50 μL of 6 ng/μL rhAKR1C4, and start the reaction by adding 50 μL of Reaction Mixture.
  4. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of Reaction Mixture.
  5. Read at an absorbance of 340 nm in kinetic mode for 5 minutes.
  6. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

     *Adjusted for Substrate Blank 
     **Using the extinction coefficient 6270 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Well:
  • rhAKR1C4: 0.3 μg
  • Sodium cholate: 1 mM
  • NADP+: 0.5 mM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Aldo-keto Reductase 1C4/AKR1C4

Aldo-keto Reductase 1C4 (AKR1C4), also known as dihydrodiol dehydrogenase 4, is a member of the aldo-keto reductase (AKR) superfamily. It catalyzes the reversible oxidoreduction of various 3 alpha ‑hydroxysteroids and prostaglandins as well as the oxidation of trans‑dihydrodiols of aromatic hydrocarbons and alicyclic alcohols (1‑4). AKR1C4 is important for steroid hormone metabolism primarily through synthesis and clearance of testosterone. Its high expression in liver and the ability to reduce xenobiotic carbonyl compounds suggests an important role in xenobiotic metabolism, as is manifested by the detoxification of the pesticide chlordecone in humans (5). AKR1C4 also plays a role in the pharmacokinetics of various drugs and prodrug activation (6, 7).

References
  1. Penning, P. M. et al. (1990) Steroids 47:221.
  2. Takikawa, H. et al. (1992) Hepatology 16:365.
  3. Binstock, J. M. et al. (1992) Biochem. Biophys. Res. Commun. 187:760.
  4. Ohara, H. et al. (1994) Biochim. Biophys. Acta. 1215:59.
  5. Winters, C. J. et al. (1990) Biochemistry 29:1080.
  6. Jin, Y. et al. (2009) J. Biol. Chem. 284:10013.
  7. Breyer-Pfaff, U. et al. (2004) J. Pharm. Pharmacol. 56:1601.

Long Name
Aldo-keto Reductase Family 1, Member C4
Entrez Gene IDs
1109 (Human)
Alternate Names
3-alpha-HSD; AKR1C4; Aldo-keto Reductase 1C4; aldo-keto reductase family 1, member C4; AldoketoReductase 1C4; C11; CDR; CHDR; DD4; DD-4; HAKRA

Product Specific Notices

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

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