ATP synthase Specific Activity Microplate Assay Kit (ab109716)


  • Product name
    ATP synthase Specific Activity Microplate Assay Kit
    See all ATP Synthase kits
  • Detection method
  • Sample type
    Cell culture supernatant, Cell culture extracts, Tissue, Tissue Extracts, Cell Lysate
  • Assay type
    Enzyme activity
  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Rat, Cow, Human
    Does not react with: Mouse
  • Product overview

    ab109716 rapid multiplexing microplate kit 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. The ATP synthase enzyme is immunocaptured within the wells of the microplate and the enzyme activity is measured by monitoring the decrease in absorbance at 340 nm. Specifically, 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. Subsequently, in these same wells, the quantity of ATP synthase is determined by adding an ATP synthase specific antibody conjugated with alkaline phosphatase. This phosphatase changes the substrate pNPP from colorless to yellow (OD 405 nm) in a time dependent manner proportional to the amount of protein captured in the wells. Included for optimal performance of the assay are buffer, detergent, antibody, substrate, lipid mix and a 96-well microplate pre-coated with monoclonal antibody.

  • 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.

  • Platform



  • 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).



This product has been referenced in:
  • Costa B  et al. TSPO ligand residence time influences human glioblastoma multiforme cell death/life balance. Apoptosis 20:383-98 (2015). Read more (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 . Read more (PubMed: 25023514) »

See all 5 Publications for this product

Customer reviews and Q&As

The capture and detection antibody used in ab109716ATP synthase Specific Activity Microplate Assay Kit are widely cross-reactive, but not tested in prokaryotes. All tested species are listed on the individual datasheets.

Having discussed ...

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1) Yes, the mitochondrial isoaltion kit is suitable for use with all kits listed.

2) Suspend the prepared mitochondria to 5.5 mg/mL as described in the attached document and follow from step 5 (adding detergent to mitochondia) to generat...

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1. I would like to know if you mean, after disrupting cell clumps by pipetting up and down, to freeze the cells "as is"? An homogenization with Teflon dounce is not necessary?

Yes resuspend cells from cell culture by pipetting and ...

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The detector antibody was raised against the ATP synthase subunit beta, mitochondrial:

We do not know the targetted subunit of the capture antibody. Our best immunocapture antibodies bind the nat...

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In the case of this product the capture binds the F1 domain – probably alpha and beta.

The detector we do know binds the beta subunit.

My colleague in our MitoSciences lab has sent me the below reply to your questions. If you need any additional information, please let me know and I will be happy to help you further.

Note this assay goes in reverse – ATP hydrolysis. Th...

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Thank you for contacting us.

Same process for converting all to molarity per amount of protein loaded into well.
Note these are all per cm and should be converted into path length for microplate which should be about 0.6cm.


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Our development scientist recommends the following units: The units for MS443: -According the protocol (change of OD505/min)/OD405 -Alternative method (change of OD505/min)/(change OD405/min) The units for MS543: -According the protocol (change of OD3...

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Both MS443 and MS543 allow determination of specific target activities, defined as ratio of target activity to target quantity. For example, to determine specific activity of COX in a well using MS443, the COX activity (in mOD550/min) divided by COX qu...

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Thank you for your enquiry. We are pleased to provide information: The best way to present the results is simply to say that the activity of ATP synthase enzyme is decreased.  Since you are not measuring ATP synthesis, saying that ATP synthesis ...

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