Aldo-keto Reductases (AKRs) are a superfamily of NAD(P)H linked oxidoreductases which contains more than 190 members. A complete listing of AKR superfamily can be found at: https:/www.med.upenn.edu/akr/. The common function of AKR superfamily is to oxidize/reduce aldehydes or ketones to their corresponding primary or secondary alcohols. Due to their broad substrate specificity, AKRs play roles in intermediary metabolism, biosynthesis and detoxification. In humans, the AKRs are not only involved in carbonyl metabolism, but they also regulate signaling through nuclear receptors which leads to chemical carcinogenesis. Many studies demonstrate increasing expression of AKRs in cancers such as lung, liver and colon cancers. Assay Genie's Aldo-keto Reductase Activity Assay Kit provides a convenient tool for sensitive detection of AKR activity in a variety of samples. In this assay, AKR acts on a general substrate and converts NADP+ to NADPH. This NADPH reacts with the AKR probe and generates color. This color is proportional to the activity of AKR in the sample. This kit can detect AKR activity as low as 0.001 mU in samples and has been validated with AKR1B10, AKR1C1 and AKR1C3. Further tests are in progress for other members of the AKR family.

Figure: (a) NADPH standard curve; (b) AKR Positive Controls (µl/assay). (c) Human tissue samples and cancer cell line (HepG2: hepatocellular carcinoma; MDA-MB: adenocarcinoma; Jurkat: acute T cell leukemia) lysates were homogenized as described in the kit protocol. Serial dilutions were tested to ensure the readings were within the linear range of the Standard Curve. AKR activities (mU/mg) were measured and calculated as in the kit protocol. Assay Genie’s AKR activity assay kit demonstrated enhanced AKR activities in the liver and liver cancer cell line lysates.