Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 4 hr
- Sample type: Cell culture extracts, Tissue Extracts
Product nameMitochondrial Aldehyde Dehydrogenase (ALDH2) Activity Assay Kit
See all Aldehyde Dehydrogenase kits
Intra-assay Sample n Mean SD CV% 1 20 5% Inter-assay Sample n Mean SD CV% 1 4 12%
Sample typeCell culture extracts, Tissue Extracts
Assay typeEnzyme activity
Range60 µg/ml - 250 µg/ml
Assay time4h 00m
Species reactivityReacts with: Mouse, Rat, Cow, Dog, Human
Mitochondrial Aldehyde Dehydrogenase (ALDH2) Activity Assay Kit ab115348 is used to determine mitochondrial aldehyde dehydrogenase activity (ALDH2) in a sample. The enzyme is captured within the wells of the microplate and activity is determined by following the production of NADH in the following ALDH2 catalyzed reaction:
acetaldehyde + NAD+ -> acid + NADH
The generation of NADH in the aldehyde dehydrogenase assay protocol is coupled to the 1:1 reduction of a reporter dye to yield a colored (yellow) reaction product whose concentration can be monitored by measuring the increase in absorbance at 450 nm (Dye molar extinction coefficient - 37000 M-1cm-1). ab115348 immunocaptures in each well only native ALDH2 from the chosen sample; this removes all other enzymes, including unrelated aldehyde dehydrogenases.
Reagent dye, Coupler and NAD+ are shipped lyophilized. Before use rehydrate by adding 0.25 mL pure water to each tube and vortex each tube thoroughly to dissolve. After hydration unused amounts of these three materials should be stored at -80°C. Store all other components store at 4°C. This kit is stable for 6 months from receipt.
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 100X acetaldehyde (2.5 M) 1 x 0.25ml 100X Coupler 1 x 0.25ml 100X NAD+ 1 x 0.25ml 100X Reagent Dye 1 x 0.25ml 10X Blocking Buffer 1 x 6ml 1X Base Buffer 1 x 24ml 20X Buffer 1 x 20ml 96 well microplate (12 strips) 1 unit Extraction Buffer (ab260490) 1 x 15ml
- Bovine Heart Mitochondria (ab110338)
- Rat liver tissue lysate - mitochondrial extract (ab110346)
- Rat heart tissue lysate - mitochondrial extract (ab110347)
- Rat brain tissue lysate - mitochondrial extract (ab110348)
- Mouse liver tissue lysate - mitochondrial extract (ab110349)
- Mouse heart tissue lysate - mitochondrial extract (ab110350)
- Mouse brain tissue lysate - mitochondrial extract (ab110351)
Determination of effect reactive species on ALDH2 activity.
The mitochondrial location of ALDH2 may increase the exposure of this enzyme to high levels of reactive oxygen and nitrogen species generated in mitochondria. ALDH2 has been shown to be specifically inactivated by nitrative stress, particularly by modification or crosslinking of an active site cysteine. To demonstrate this rat liver homogenate was exposed in vitro to a dilution series of peroxynitirite at concentrations - 0, 25, 50, 100, 200, 400, 800, 1600 µM. After exposure the samples were extracted, diluted to 0.25 mg/mL and loaded into microplate wells in duplicate. After following the assay protocol as described above, the activity of each sample was measured and graphed against peroxynitrite exposure concentration. The ALDH2 activity after exposure was reduced with an IC50approximately 210 µM.
Reference Sample graph
Reference sample graph
ab115348 has been referenced in 16 publications.
- Matsumura Y et al. Systemic Adeno-Associated Virus-Mediated Gene Therapy Prevents the Multiorgan Disorders Associated with Aldehyde Dehydrogenase 2 Deficiency and Chronic Ethanol Ingestion. Hum Gene Ther 31:163-182 (2020). PubMed: 31801381
- Zhao L et al. Aldehyde dehydrogenase 2 protects cardiomyocytes against lipotoxicity via the AKT/glycogen synthase kinase 3 beta pathways. Biochem Biophys Res Commun 525:360-365 (2020). PubMed: 32089266
- Katary M & Abdel-Rahman AA Alcohol suppresses cardiovascular diurnal variations in male normotensive rats: Role of reduced PER2 expression and CYP2E1 hyperactivity in the heart. Alcohol 89:27-36 (2020). PubMed: 32777474
- Guillot A et al. Targeting liver aldehyde dehydrogenase-2 prevents heavy but not moderate alcohol drinking. Proc Natl Acad Sci U S A 116:25974-25981 (2019). PubMed: 31792171
- Cao R et al. ALDH2 Overexpression Alleviates High Glucose-Induced Cardiotoxicity by Inhibiting NLRP3 Inflammasome Activation. J Diabetes Res 2019:4857921 (2019). PubMed: 31871948