AKR1B1 蛋白, Human (His)

The AKR1B1 protein uses NADPH to reduce various carbonyl compounds to alcohols. AKR1B1 Protein, Human (His) is the recombinant human-derived AKR1B1 protein, expressed by E. coli , with N-10*His labeled tag.

AKR1B1 是一种多功能酶，可催化多种含羰基化合物依赖 NADPH 还原为其各自的醇。它对内源代谢物表现出酶活性，包括芳香族和脂肪族醛、酮、单糖、胆汁酸和异生物质底物。作为多元醇途径中的关键酶，AKR1B1 通过催化葡萄糖还原为山梨醇，在高血糖中发挥着关键作用。该酶还可以减少类固醇及其衍生物、前列腺素，并对视网膜表现出低活性。此外，AKR1B1 还参与膳食和脂质衍生的不饱和羰基化合物的解毒，例如巴豆醛、4-羟基壬烯醛、反式-2-己烯醛、反式-2,4-己二醛及其谷胱甘肽缀合羰基化合物 (GS-羰基化合物) 。值得注意的是，它催化磷脂酰胆碱和磷脂酰乙醇酰胺氧化产生的各种磷脂醛的还原，进一步强调了其广泛的底物特异性及其在细胞解毒过程中的不可或缺的作用。

Measured by its ability to catalyzes glucuronic acid decomposition in the presence of NADPH. The specific activity is 132.9 U/mg.

Materials
AKR1B1 Protein, Human (His) (HY-P75521): Dissolve in deionized water to a concentration of 100 μg/mL.
Substrate: Glucuronic acid. Accurately weigh 15.520 mg of powder and dissolve in 1 mL of deionized water to a concentration of 80 mM.
Standard: NADPH. Accurately weigh 3.33 mg of solid powder and dissolve in 1 mL of deionized water to a concentration of 4 mM.

Procedure
1. Dilute NADPH to the following concentrations: 0, 0.015625, 0.03125, 0.0625, 0.125, 0.25 mM. Add 200 μL of each concentration to a 96-well plate in triplicate.
2. Measure the absorbance at 340 nm using a microplate reader. Plot a standard curve with the amount of NADPH as the x-axis and the OD value as the y-axis.
3. Dilute the substrate to a concentration of 8 mM for later use.
4. Dilute Human AKR1B1 to 0.2 μg/mL. Mix 50 μL of the substrate with 50 μL of the protein at each concentration, and pre-add to a 96-well plate in triplicate.
5. Dilute 4 mM NADPH to 0.4 mM. Initiate the reaction by adding 100 μL of 0.4 mM NADPH to each well.
6. Zero the microplate reader using the blank group OD value. Measure the absorbance at 340 nm in kinetic mode for 10 min.
7. Define one enzyme unit as the consumption of 1 μmol of NADPH per minute, denoted as U. Calculate the specific enzyme activity.

Human

E. coli

N-10*His

P15121 (M1-F316)

231  [NCBI]

Full Length

MASRLLLNNGAKMPILGLGTWKSPPGQVTEAVKVAIDVGYRHIDCAHVYQNENEVGVAIQEKLREQVVKREELFIVSKLWCTYHEKGLVKGACQKTLSDLKLDYLDLYLIHWPTGFKPGKEFFPLDESGNVVPSDTNILDTWAAMEELVDEGLVKAIGISNFNHLQVEMILNKPGLKYKPAVNQIECHPYLTQEKLIQYCQSKGIVVTAYSPLGSPDRPWAKPEDPSLLEDPRIKAIAAKHNKTTAQVLIRFPMQRNLVVIPKSVTPERIAENFKVFDFELSSQDMTTLLSYNRNWRVCALLSCTSHKDYPFHEEF

Approximately 36 kDa

• 
  Greater than 95% as determined by reducing SDS-PAGE.

无菌粉末.

Lyophilized from a 0.2 μm filtered solution of PB, 150 mM NaCl, pH 7.4.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.