Mitochondrial Aldehyde Dehydrogenase (ALDH2) Activity Assay Kit (ab115348)
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
Overview
-
Product name
Mitochondrial Aldehyde Dehydrogenase (ALDH2) Activity Assay Kit
See all Aldehyde Dehydrogenase kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% 1 20 5% Inter-assay Sample n Mean SD CV% 1 4 12% -
Sample type
Cell culture extracts, Tissue Extracts -
Assay type
Enzyme activity -
Range
60 µg/ml - 250 µg/ml -
Assay time
4h 00m -
Species reactivity
Reacts with: Mouse, Rat, Cow, Dog, Human -
Product overview
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.
-
Notes
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.
-
Platform
Microplate reader
Properties
-
Storage instructions
Store 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 -
Research areas
Images
-
Functional Studies - Mitochondrial Aldehyde Dehydrogenase (ALDH2) Activity Assay Kit (ab115348)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. -
Functional Studies - Mitochondrial Aldehyde Dehydrogenase (ALDH2) Activity Assay Kit (ab115348)Reference Sample graph -
Functional Studies - Mitochondrial Aldehyde Dehydrogenase (ALDH2) Activity Assay Kit (ab115348)Reference sample graph