ATP synthase Specific Activity Microplate Assay Kit (ab109716)
Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Cell culture extracts, Cell culture supernatant, Cell Lysate, Tissue, Tissue Extracts
Product nameATP synthase Specific Activity Microplate Assay Kit
See all ATP Synthase kits
Sample typeCell culture supernatant, Cell culture extracts, Tissue, Tissue Extracts, Cell Lysate
Assay typeEnzyme activity
Species reactivityReacts with: Rat, Cow, Human
Does not react with: Mouse
ATP Synthase Activity Assay Kit ab109716 is used to determine both the activity and quantity of ATP synthase (Complex V) in a human or rat sample. The ratio of the two measurements represents the enzyme's specific activity.
In the ATP synthase assay protocol, the ATP synthase enzyme is immunocaptured within the wells of a microplate. The conversion of ATP to ADP by ATP synthase is coupled to the oxidative reaction of NADH to NAD+. The formation of NAD+ results in a decrease in absorbance at 340 nm which is proportional to the amount of enzyme activity.
Subsequently, the quantity of ATP synthase is determined by adding an anti-ATP synthase antibody conjugated with alkaline phosphatase. Alkaline phosphatase acts on the substrate pNPP to form a yellow color (OD 405 nm) proportional to the amount of ATP synthase captured in the wells.
ATP synthase assay protocol summary:
- add samples to wells and incubate for 3 hrs to capture enzyme
- add lipid mix and incubate for 45 min
- add reaction mix
- analyze enzyme activity with microplate reader for 1-2 hrs at 1 min intervals
- empty wells and incubate with solution A for 1 hr
- add solution B and incubate for 1 hr
- add development solution
- analyze enzyme quantity with microplate reader for 30 min at 1 min intervals
Store Buffer, Detergent, Detector Antibody, AP Label, AP Buffer, and the microplate at 4°C. Store the Reagent Mix, AP Development Reagent and Lipid Mix at -20°C. For multiple experiments, the Reagent Mix and AP Development Reagent may be aliquoted and stored at -20°C, or at -80°C for long term storage.
Storage instructionsPlease refer to protocols.
Components Identifier 96 tests AP ATP synthase Detector antibody 1 x 1ml AP Buffer Tube 2 1 x 20ml AP Development Reagent 1 x 400µl AP label 2500 X Tube B 1 x 12µl ATP synthase precoated Microtiter plate 1 unit Buffer Tube 1 1 x 15ml Detergent 1 x 1ml Phospholipids 1 x 6ml Reagent Mix 1 x 20ml
- Complex V
- F1F0 ATP synthase
- SwissProt: P25705 Human
- SwissProt: P10719 Rat
By measuring both quantity and activity of ATP synthase in a sample, the ratio of the two parameters can be calculated. This is the relative specific activity and should be compared to the normal or control.
Examples of quantity/load relationships for cultured liver cell lysate, fibroblast lysate, and rat liver mitochondria.
Functional study using ATP synthase Specific Activity Microplate Assay Kit (ab109716) measuring enzyme quantity.
Functional study using ATP synthase Specific Activity Microplate Assay Kit (ab109716).
Examples of activity/load relationships for cultured cell lysate, rat liver mitochondria, and bovine heart mitochondria.
Functional study using ATP synthase Specific Activity Microplate Assay Kit (ab109716) measuring enzyme activity.
Functional study using ATP synthase Specific Activity Microplate Assay Kit (ab109716)
Principle of ATP synthase Specific Activity Microplate Assay Kit (ab109716).
Datasheets and documents
ab109716 has been referenced in 9 publications.
- Puca F et al. Medium-Chain Acyl-CoA Dehydrogenase Protects Mitochondria from Lipid Peroxidation in Glioblastoma. Cancer Discov 11:2904-2923 (2021). PubMed: 34039636
- Yao PJ et al. Mitochondrial Electron Transport Chain Protein Abnormalities Detected in Plasma Extracellular Vesicles in Alzheimer's Disease. Biomedicines 9:N/A (2021). PubMed: 34829816
- Chen R et al. Parkinson's disease protein DJ-1 regulates ATP synthase protein components to increase neuronal process outgrowth. Cell Death Dis 10:469 (2019). PubMed: 31197129
- Tran L et al. Mitochondrial ATP synthase ß-subunit production rate and ATP synthase specific activity are reduced in skeletal muscle of humans with obesity. Exp Physiol 104:126-135 (2019). PubMed: 30362197
- Costa B et al. TSPO ligand residence time influences human glioblastoma multiforme cell death/life balance. Apoptosis 20:383-98 (2015). PubMed: 25413799
- Rahman M et al. Drosophila Sirt2/mammalian SIRT3 deacetylates ATP synthase ß and regulates complex V activity. J Cell Biol 206:289-305 (2014). Functional Studies . PubMed: 25023514
- Maniam S et al. Cofactor Strap regulates oxidative phosphorylation and mitochondrial p53 activity through ATP synthase. Cell Death Differ N/A:N/A (2014). Functional Studies . PubMed: 25168243
- Fan X et al. Dynamic proteomics of nucleus accumbens in response to acute psychological stress in environmentally enriched and isolated rats. PLoS One 8:e73689 (2013). Functional Studies . PubMed: 24040027
- Ito E et al. Potential use of cetrimonium bromide as an apoptosis-promoting anticancer agent for head and neck cancer. Mol Pharmacol 76:969-83 (2009). PubMed: 19654225