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

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

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 #: O60218
• 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

• Dilutions:
  • Enzyme Activity
• 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

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 10^12 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.

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

Average Rating: 5

(Based on 1 review)

reviewed by: Verified Customer Enzymatic activity in vitro 10/10/2018

Summary

• Application: Enzymatic activity in vitro
• Lot: DCB00216091

Bioinformatics

• Gene Symbol: AKR1B10
• Uniprot:
  • O60218()