Aldo-Keto reductase family1, member B1, 1-316aa, Recombinant, Human

Aldose reductase (AKR1B1) is a member of the aldo-keto reductase (AKR) superfamily and catalyzes the NADPH-dependent reduction of a wide variety of carbonyl-containing compounds to their corresponding alcohols. This protein is implicated in the development of diabetic complications by catalyzing the reduction of glucose to sorbitol. Source: Human Aldose reductase (AKR1B1) protein was expressed in E. coli and purified by using conventional chromatography techniques. Specific Activity: ~0.2-0.9 units/mg protein. Enzymatic activity was confirmed by measuring the amount of enzyme catalyzing the oxidation of 1 micromole NADPH per minute at 25°C. , Activity Assay: 1. Prepare a 750ul reaction mix into a suitable container. The final concentrations of the reaction mix components should be: 0.1M sodium phosphate (pH7.0) and 0.3mM NADPH. , 2. Add 50ul of recombinant AKR1B1 solution with various concentrations (2.5ug, 5ug, 10ug) in 750ul reaction buffer. , 3. Mix by inversion and Incubate at 25°C for 2.5 minutes., 4. Add 200ul of 50 mM DL-glyceraldehyde substrate and immediately mix by inversion., 5. Record the increase in A340nm for 3 minutes. AA Sequence: MASRLLLNNG AKMPILGLGT WKSPPGQVTE AVKVAIDVGY RHIDCAHVYQ NENEVGVAIQ EKLREQVVKR EELFIVSKLW CTYHEKGLVK GACQKTLSDL KLDYLDLYLI HWPTGFKPGK EFFPLDESGN VVPSDTNILD TWAAMEELVD EGLVKAIGIS NFNHLQVEMI LNKPGLKYKP , AVNQIECHPY LTQEKLIQYC QSKGIVVTAY SPLGSPDRPW AKPEDPSLLE DPRIKAIAAK HNKTTAQVLI RFPMQRNLVV IPKSVTPERI AENFKVFDFE LSSQDMTTLL SYNRNWRVCA LLSCTSHKDY PFHEEF , Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.