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Recombinant Human Aldo-keto Reductase 1C3/AKR1C3 Protein, CF

Catalog #: 7678-DH
1 Citation 1 Review Datasheet / COA / SDS
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7678-DH-020
R&D Systems Recombinant 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 1C3/AKR1C3 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 ability to catalyze the reduction of phenanthrenequinone. The specific activity is >115 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived human Aldo-keto Reductase 1C3/AKR1C3 protein
Asp2-Tyr323, with an N-terminal Met and 6-His tag
Accession #
P42330
N-terminal Sequence
Analysis
Met
Predicted Molecular Mass
38 kDa
SDS-PAGE
36-37 kDa, reducing conditions

Product Datasheets

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7678-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.

7678-DH

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, Brij and Glycerol.
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: 100 mM Sodium Phosphate, pH 6.0
  • Recombinant Human Aldo‑keto Reductase 1C3/AKR1C3 (rhAKR1C3) (Catalog # 7678-DH)
  • 9,10-Phenanthrenequinone (PQ) (Sigma, Catalog # 156507), 5 mM stock in N,N-Dimethylformamide
  • beta -Nicotinamide adenine dinucleotide phosphate reduced tetrasodium salt ( beta -NADPH) (Sigma, Catalog # N7505), 10 mM in deionized water
  • UV Plate (Costar, Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhAKR1C3 to 20 μg/mL in Assay Buffer.
  2. Prepare a Reaction Mixture containing 40 μM PQ and 400 μM beta -NADPH in Assay Buffer.
  3. In a plate, load 50 μL of 20 μg/mL rhAKR1C3, 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.
  4. Read at an absorbance of 340 nm in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x -1 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:
  • rhAKR1C3: 1 μg
  • PQ: 20 μM
  • beta -NADPH: 200 μM
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 1C3/AKR1C3

AKR1C3 is a member of the aldo-keto reductase (AKR) superfamily. It catalyzes oxidation/reduction reactions at the 3-alpha, 20-alpha, and 17-beta positions of steroids (1). It is also known as Prostaglandin F synthase as it reduces prostaglandin D2 to F2, and therefore may play a role in allergic conditions such as asthma (2). Elevated expression of AKR1C3 in endometrium that results in enhanced estrogen action may lead to endometrial cancer (3). It is also up‑regulated in squamous cell carcinoma of head and neck (4). AKR1C3 is found to be a novel suppressor of cell differentiation that provides a plausible target for the non-cyclooxygenase-dependent antineoplastic actions of nonsteroidal anti-inflammatory drugs (5).

References
  1. Penning, T. M. et al. (2000) Biochem J. 351:67.
  2. Suzuki-Yamamoto, T. et al. (1999) FEBS Lett. 462:335.
  3. Rizner, T. L. et al. (2006) Mol. Cell. Endocrinol. 248:126.
  4. Li, S. et al. (2004) Br. J. Cancer 90:1093.
  5. Desmond, J. C. et al. (2003) Cancer Res. 63:505.
Long Name
Aldo-keto Reductase Family 1, Member C3
Entrez Gene IDs
8644 (Human); 105349 (Mouse); 171516 (Rat)
Alternate Names
17-beta-hydroxysteroid dehydrogenase type 5; 3-alpha-HSD type 2; 3-alpha-hydroxysteroid dehydrogenase type 2; AKR1C3; Aldo-keto Reductase 1C3; aldo-keto reductase family 1 member C3; aldo-keto reductase family 1, member C3 (3-alpha hydroxysteroid dehydrogenase, type II); AldoketoReductase 1C3; Chlordecone reductase homolog HAKRb; DD3; DD-3; DD317-beta-HSD 5; DDH1; DDX; Dihydrodiol dehydrogenase 3; Dihydrodiol dehydrogenase type I; dihydrodiol dehydrogenase X; EC 1.1.1; EC 1.1.1.112,3-alpha-HSD type II, brain; EC 1.1.1.188; EC 1.1.1.213; EC 1.1.1.63; EC 1.1.1.64; EC 1.3.1.20; HA1753; HAKRB; HAKRBtype IIb 3-alpha hydroxysteroid dehydrogenase; HAKRe; HSD17B5; hydroxysteroid (17-beta) dehydrogenase 5; Indanol dehydrogenase; KIAA0119hluPGFS; PGFS; Prostaglandin F synthase; Testosterone 17-beta-dehydrogenase 5; Trans-1,2-dihydrobenzene-1,2-diol dehydrogenase

Product Specific Notices

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

Citation for Recombinant Human Aldo-keto Reductase 1C3/AKR1C3 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.

1 Citation: Showing 1 - 1

  1. Adipocytes Sequester and Metabolize the Chemotherapeutic Daunorubicin
    Authors: X Sheng, JH Parmentier, J Tucci, H Pei, O Cortez-Tol, CM Dieli-Conw, MJ Oberley, M Neely, E Orgel, SG Louie, SD Mittelman
    Mol. Cancer Res., 2017-11-08;0(0):.
    Applications: Bioassay

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Recombinant Human Aldo-keto Reductase 1C3/AKR1C3 Protein, CF
By Anonymous on 11/16/2018
Application: Enzymatic activity in vitro
Reason for Rating: Time course vs OD
Enzymatic activity in vitro Recombinant Human Aldo-keto Reductase 1C3/AKR1C3 Protein, CF 7678-DH