AKR1B1 Protein, Human (His)
Based on 1 Customer Validation
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.
- Species: Human
- Source: E. coli
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Storage: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.
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Biological Activity
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Technical Parameters
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Product Properties
- Documentation
- Help & FAQs
Biological Activity
Description
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.
Background
AKR1B1, a versatile enzyme, catalyzes the NADPH-dependent reduction of a diverse array of carbonyl-containing compounds to their respective alcohols. It exhibits enzymatic activity towards endogenous metabolites, including aromatic and aliphatic aldehydes, ketones, monosaccharides, bile acids, and xenobiotics substrates. As a key enzyme in the polyol pathway, AKR1B1 plays a critical role in hyperglycemia by catalyzing the reduction of glucose to sorbitol. The enzyme also reduces steroids and their derivatives, prostaglandins, and displays low activity toward retinals. Additionally, AKR1B1 is involved in the detoxification of dietary and lipid-derived unsaturated carbonyls, such as crotonaldehyde, 4-hydroxynonenal, trans-2-hexenal, trans-2,4-hexadienal, and their glutathione-conjugate carbonyls (GS-carbonyls). Notably, it catalyzes the reduction of various phospholipid aldehydes generated from the oxidation of phosphatidylcholine and phosphatidylethanolamides, further emphasizing its broad substrate specificity and its integral role in cellular detoxification processes.
Verified Bioactivity
Measured by its ability to catalyzes glucuronic acid decomposition in the presence of NADPH. The specific activity is 132.9 U/mg.
MCE Validation Data
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Purity - SDS-PAGE
Purity - SDS-PAGE
Assay Procedure
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.
Specific Activity (U/mg) = | ΔnNADPH (μmol) |
| Reaction time (min) x amount of enzyme (mg) |
Per Well:
Human AKR1B1: 0.01 μg
Substrate: 2 mM
NADPH: 0.2 mM
Technical Parameters
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Species Human
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Source E. coli
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Tag N-10*His
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Accession
P15121 (M1-F316)
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Molecular Construction
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N-term
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10*His
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AKR1B1 (M1-F316)
Accession # P15121 -
C-term
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Protein Length
Full Length
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Synonyms
AKR1B1; ALR2; Prev. ALDR1; Lii5-2 CTCL Tumor Antigen; Aldose Reductase; Low Km Aldose Reductase; AR; Aldehyde Reductase 1; Aldo-Keto Reductase Family 1 Member B1; ADR; Aldehyde Reductase; Aldo-Keto Reductase Family 1 Member B; Aldo-Keto Reductase Family 1
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AA Sequence
MASRLLLNNGAKMPILGLGTWKSPPGQVTEAVKVAIDVGYRHIDCAHVYQNENEVGVAIQEKLREQVVKREELFIVSKLWCTYHEKGLVKGACQKTLSDLKLDYLDLYLIHWPTGFKPGKEFFPLDESGNVVPSDTNILDTWAAMEELVDEGLVKAIGISNFNHLQVEMILNKPGLKYKPAVNQIECHPYLTQEKLIQYCQSKGIVVTAYSPLGSPDRPWAKPEDPSLLEDPRIKAIAAKHNKTTAQVLIRFPMQRNLVVIPKSVTPERIAENFKVFDFELSSQDMTTLLSYNRNWRVCALLSCTSHKDYPFHEEF
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Molecular Weight
Approximately 36 kDa, based on SDS-PAGE under reducing conditions.
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Purity
≥ 95%, as determined by reducing SDS-PAGE.
Product Properties
Lyophilized powder.
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.
Documentation
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Data Sheet (237 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Handling Instructions (2659 KB)
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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