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

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Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
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Recombinant Human Aldo-keto Reductase 1C3/AKR1C3 Protein, CF Summary

Details of Functionality
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 #
N-terminal Sequence
Met
Protein/Peptide Type
Recombinant Enzymes
Gene
AKR1C3
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
38 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
36-37 kDa, reducing conditions
Reviewed Applications
Read 1 Review rated 5
using
7678-DH in the following application:
Publications
Read Publication using 7678-DH.

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in Tris, NaCl, Brij and Glycerol.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Assay Procedure
  • 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

Notes

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

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

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

  • 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

Background

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).
  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.

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Publications for Aldo-keto Reductase 1C3/AKR1C3 (7678-DH)(1)

We have publications tested in 1 application: Bioassay.


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Review for Aldo-keto Reductase 1C3/AKR1C3 (7678-DH) (1) 51

Average Rating: 5
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Enzymatic activity in vitro Aldo-keto Reductase 1C3/AKR1C3 7678-DH
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ApplicationEnzymatic activity in vitro
LotDCJJ0117041

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Bioinformatics

Gene Symbol AKR1C3
Uniprot