Biological Information

Background Information:

Aldose reductase, a member of the monomer NADPH-dependent aldo-keto reductase, is a rate-limiting enzyme in the polyol pathway which catalyzes the reduction of various aldehydes. This includes reduction of the aldehyde form of glucose to its corresponding sugar alcohol sorbitol. Accumulation of sorbitol has been reported in the lens, nerve, kidney, and retina of diabetic animals. Large amounts of sorbitol cause osmotic disruption, which may be one of the etiologic factors in the pathogenesis of some diabetic complications.

Family:

Steroid Metabolism

Sub Family:

Oxidoreductase

Class:

Lipid metabolism

Protein Name:

Aldose Reductase

Uniprot Number:

P15121

Protein Aliases:

aldehyde reductase

Gene Name:

AKR1B1

Gene ID:

231

Gene Aliases:

AR

Accession Number:

NM_001628.4

Organism:

Human

Construct Details:

Full length, recombinant

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Antagonist

Detection Method:

Spectrophotometry

Platform:

Plate reader

Measured Response:

Absorbance

Testing Information

Procedure Summary:

Human recombinant aldose reductase expressed in E. coli is used. Test compound and/or vehicle is preincubated with 5 µg/ml enzyme and 0.2 mM NADPH in phosphate buffer pH 6.2 for 15 minutes at 25°C. The reaction is initiated by addition of 250 µM DL-glyceraldehyde for another 20-minute incubation period. Determination of the amount of NADPH remained is read spectrophotometrically at 340 nm. Compounds are screened at 10 µM.

Substrate:

250 µM DL-Glyceraldehyde

Incubation:

20 min at 25°C

Control Inhibitor:

Quercitrin

Criteria For Significance:

≥ 50% of max stimulation or inhibition

Test Concentration / Dose:

Suggested concentration for initial testing is 10 uM

Test Sample Requirements:

Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).

Minimum Order Quantity:

2

Turnaround Time

Standard:

15 Business Days

Reference Compound Data

PubChem ID

5280459

Name

Quercitrin

Measurement

3.6μM - IC50

Clinical Relevance

Therapeutic Area:

Metabolic Diseases

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

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