Recombinant Human Aldo-keto Reductase 1B10/AKR1B10, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human Aldo-keto Reductase 1B10/AKR1B10, CF Summary

Details of Functionality
Measured by the disappearance of NADPH during the reducation of 4-nitrobenzaldehyde. The specific activity is >1,000 pmol/min/μg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Aldo-keto Reductase 1B10/AKR1B10 protein
Ala2-Tyr316, with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Inconclusive result, Met predicted. Protein identity confirmed by MS analysis of tryptic fragments
Protein/Peptide Type
Recombinant Enzymes
Gene
AKR1B10
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Note
<0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
37 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
37-38 kDa, reducing conditions
Reviewed Applications
Read 1 Review rated 5
using
7529-DH in the following application:

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, DTT 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: 50 mM Sodium Acetate, 0.1 M NaCl, pH 5.0
  • Recombinant Human Aldo‑keto Reductase 1B10/AKR1B10 (rhAKR1B10) (Catalog # 7529-DH)
  • beta -Nicotinamide adenine dinucleotide phosphate reduced, tetrasodium salt ( beta -NADPH) (Sigma, Catalog # N7505), 10 mM in deionized water
  • 4-nitro-benzaldehyde (Fluka, Catalog # 72800), 200 mM in DMSO
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhAKR1B10 to 4 ng/μL in Assay Buffer.
  2. Prepare a Reaction Mixture containing 2 mM 4-nitro-benzaldehyde and 400 μM beta -NADPH in Assay Buffer.
  3. In a plate, load 50 μL of 4 ng/μL rhAKR1B10, 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 -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:
  • rhAKR1B10: 0.2 μg
  • 4-nitro-benzaldehyde: 1 mM
  • 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 1B10/AKR1B10, CF

  • AKR1B10
  • AKR1B11
  • AKR1B12
  • Aldo-keto Reductase 1B10
  • aldo-keto reductase family 1 member B10
  • aldo-keto reductase family 1, member B10 (aldose reductase)
  • aldo-keto reductase family 1, member B11 (aldose reductase-like)
  • AldoketoReductase 1B10
  • aldose reductase-like 1
  • aldose reductase-like peptide
  • Aldose reductase-like
  • Aldose reductase-related protein
  • ALDRLn
  • ARL1
  • ARL-1SI reductase
  • ARP
  • EC 1.1.1
  • EC 1.1.1.-
  • EC 1.1.1.21
  • hARP
  • HIS
  • HSI
  • MGC14103
  • Small intestine reductase

Background

Aldo-keto reductase family 1 member B10 (AKR1B10), also known as ARL-1 or aldose reductase related protein, is a cytosolic, NADPH-dependent oxidoreductase that reduces variety of aromatic and aliphatic aldehydes, dicarbonyl compounds, and some drug ketones (1). It is related to AKR1B1 in amino acid sequence and tertiary structure. However, its substrate specificity differs from AKR1B1 in its inability to reduce sugars and prostaglandin H2, and in its high catalytic efficiency for retinals, isoprenyl aldehydes, and cytotoxic aldehydes (2). It is primarily observed in the human colon, small intestine, and adrenal gland, with a low level in liver. Its up‑regulated expression is detected in hepatocellular carcinoma, cervical cancer, lung squamous cell carcinoma, and lung adenocarcinoma in smokers (3). Thus, it is considered as a potential diagnostic and/or prognostic marker in carcinomas and serum. The functional studies of this enzyme suggest that the up‑regulation of AKR1B10 is related to cell survival by reducing chemo-agents or metabolites in cells to less toxic reduced forms (4). Its inhibition may be a potential target for reducing the progression of cancers (5).
  1. Endo, S. et al. (2009) Arch. Biochem. Biophys. 487:1.
  2. Gallego, O. et al. (2007) Proc. Natl. Acad. Sci. USA 104:20764.
  3. Matsunaga, T. et al. (2012) Front. Pharmacol. 3:1.
  4. Wang, C. et al. (2009) J. Biol. Chem. 284:26742.
  5. Matsunaga, T. et al. (2011) Anticancer Drugs 22:402.

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Review for Aldo-keto Reductase 1B10/AKR1B10 (7529-DH) (1) 51

Average Rating: 5
(Based on 1 review)

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reviewed by:
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10/10/2018
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Summary

LotDCB00216091

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Gene Symbol AKR1B10
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