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

Catalog #: 6957-DH Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
6957-DH-020

R&D Systems Recombinant Proteins and Enzymes

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All Aldo-keto Reductase 1C4/AKR1C4 Proteins and Enzymes

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

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

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

Dilute rhAKR1C4 to 6 ng/μL in Assay Buffer.
Prepare a Reaction Mixture containing 2 mM sodium cholate and 1 mMNADP+ in Assay Buffer.
In a plate, load 50 μL of 6 ng/μL rhAKR1C4, and start the reaction by adding 50 μL of Reaction Mixture.
Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of Reaction Mixture.
Read at an absorbance of 340 nm in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (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

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