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
Overview
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Product name
ATP synthase Specific Activity Microplate Assay Kit
See all ATP Synthase kits -
Detection method
Colorimetric -
Sample type
Cell culture supernatant, Cell culture extracts, Tissue, Tissue Extracts, Cell Lysate -
Assay type
Enzyme activity -
Species reactivity
Reacts with: Rat, Cow, Human
Does not react with: Mouse -
Product overview
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
- wash
- 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
- wash
- add solution B and incubate for 1 hr
- wash
- add development solution
- analyze enzyme quantity with microplate reader for 30 min at 1 min intervals -
Notes
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.
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Platform
Microplate reader
Properties
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Storage instructions
Please 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 -
Research areas
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Alternative names
- Complex V
- F1F0
- F1F0 ATP synthase
see all -
Database links
- SwissProt: P25705 Human
- SwissProt: P10719 Rat
Images
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Functional Studies - ATP synthase Specific Activity Microplate Assay Kit (ab109716)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.
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Functional Studies - ATP synthase Specific Activity Microplate Assay Kit (ab109716)Examples of quantity/load relationships for cultured liver cell lysate, fibroblast lysate, and rat liver mitochondria. -
Functional Studies - ATP synthase Specific Activity Microplate Assay Kit (ab109716)Functional study using ATP synthase Specific Activity Microplate Assay Kit (ab109716) measuring enzyme quantity. -
Functional Studies - ATP synthase Specific Activity Microplate Assay Kit (ab109716)Functional study using ATP synthase Specific Activity Microplate Assay Kit (ab109716). -
Functional Studies - 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 Studies - ATP synthase Specific Activity Microplate Assay Kit (ab109716)Functional study using ATP synthase Specific Activity Microplate Assay Kit (ab109716) measuring enzyme activity. -
Functional Studies - ATP synthase Specific Activity Microplate Assay Kit (ab109716)Functional study using ATP synthase Specific Activity Microplate Assay Kit (ab109716) -
Functional Studies - ATP synthase Specific Activity Microplate Assay Kit (ab109716)Principle of ATP synthase Specific Activity Microplate Assay Kit (ab109716).
