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

Name: Recombinant Human Aldo-keto Reductase 1B10/AKR1B10, CF
Brand: R&D Systems
SKU: 7529-DH
Rating: 5 (1 reviews)

Catalog #: 7529-DH 1 Review Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
7529-DH-020

R&D Systems Recombinant Proteins and Enzymes

1 Image

All Aldo-keto Reductase 1B10/AKR1B10 Proteins and Enzymes

Product Details

FAQs

Reviews (1)

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

Recombinant Human Aldo-keto Reductase 1B10/AKR1B10, 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

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

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
Analysis

Inconclusive result, Met predicted. Protein identity confirmed by MS analysis of tryptic fragments

Predicted Molecular Mass

37 kDa

SDS-PAGE

37-38 kDa, reducing conditions

Product Datasheets

7529-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.

7529-DH

Formulation	Supplied as a 0.2 μm filtered solution in Tris, NaCl, DTT 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: 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

Dilute rhAKR1B10 to 4 ng/μL in Assay Buffer.
Prepare a Reaction Mixture containing 2 mM 4-nitro-benzaldehyde and 400 μM beta -NADPH in Assay Buffer.
In a plate, load 50 μL of 4 ng/μL rhAKR1B10, 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 -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

Reconstitution Calculator

Background: Aldo-keto Reductase 1B10/AKR1B10

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).

References

Endo, S. et al. (2009) Arch. Biochem. Biophys. 487:1.
Gallego, O. et al. (2007) Proc. Natl. Acad. Sci. USA 104:20764.
Matsunaga, T. et al. (2012) Front. Pharmacol. 3:1.
Wang, C. et al. (2009) J. Biol. Chem. 284:26742.
Matsunaga, T. et al. (2011) Anticancer Drugs 22:402.

Long Name

Aldo-keto Reductase Family 1, Member B10 [Aldose Reductase]

Entrez Gene IDs

57016 (Human); 67861 (Mouse); 296972 (Rat)

Alternate Names

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