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 9 publications.
- Laitakari A et al. Systemic inactivation of hypoxia-inducible factor prolyl 4-hydroxylase 2 in mice protects from alcohol-induced fatty liver disease. Redox Biol 22:101145 (2019). PubMed: 30802717
- Fang T et al. Alterations in necroptosis during ALDH2-mediated protection against high glucose-induced H9c2 cardiac cell injury. Mol Med Rep 18:2807-2815 (2018). PubMed: 30015964
- Panisello-Roselló A et al. Aldehyde Dehydrogenase 2 (ALDH2) in Rat Fatty Liver Cold Ischemia Injury. Int J Mol Sci 19:N/A (2018). PubMed: 30131474
- Choi Y et al. Preventive effects of indole-3-carbinol against alcohol-induced liver injury in mice via antioxidant, anti-inflammatory, and anti-apoptotic mechanisms: Role of gut-liver-adipose tissue axis. J Nutr Biochem 55:12-25 (2018). PubMed: 29331880
- Gu X et al. Effect of ALDH2 on High Glucose-Induced Cardiac Fibroblast Oxidative Stress, Apoptosis, and Fibrosis. Oxid Med Cell Longev 2017:9257967 (2017). PubMed: 29129988
- Jin S et al. ALDH2(E487K) mutation increases protein turnover and promotes murine hepatocarcinogenesis. Proc Natl Acad Sci U S A 112:9088-93 (2015). Functional Studies . PubMed: 26150517
- Cummins TD et al. Metabolic remodeling of white adipose tissue in obesity. Am J Physiol Endocrinol Metab 307:E262-77 (2014). PubMed: 24918202
- Paneni F et al. Deletion of the activated protein-1 transcription factor JunD induces oxidative stress and accelerates age-related endothelial dysfunction. Circulation 127:1229-40, e1-21 (2013). Mouse . PubMed: 23410942
- Khojasteh SC et al. Characterization of rat liver proteins adducted by reactive metabolites of menthofuran. Chem Res Toxicol 25:2301-9 (2012). Rat . PubMed: 23106769