Description
Key facts
Detection method:Colorimetric,
Sample types:Tissue Lysate, Cell Lysate,
Assay type:Enzyme activity (quantitative),
Range:1 - 5 nmol/well,
Assay Platform:Microplate reader
Product details:
ATPase Assay Kit (Colorimetric) (ab234055) provides a quick and easy method for monitoring ATPase activity in cell and tissue lysates.
ATPase assay principle
In the ATPase assay protocol, ATPase hydrolyzes ATP releasing ADP and a free phosphate ion, and through linked reactions, a strong, stable chromophore is generated (OD 650 nm).
ATPases catalyzes the decomposition of ATP into ADP and a free phosphate ion. This assay can be used in the study of the many classes of ATPases including Na+ /K+ -ATPase, H + /K+ -ATPase, Ca2+ -ATPase, etc.
ATPase assay protocol summary
- Prepare all samples, controls and standards.
- Add Reaction Mix to each well containing Positive Control, Reagent Control and test samples.
- Incubate at 25°C for 30 min. Do not add Reaction Mix to the Standards.
- Add ATPase Assay Developer to all standards, ATPase Positive Control, Test Samples and Sample Background Controls.
- Incubate at 25°C for 30 minutes.
- Measure OD at 650 nm in Endpoint mode.
Other Notes
This product was previously called K417 Biovision ATPase Activity Assay Kit (Colorimetric). Biovision was acquired by Abcam in 2021.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
ATPase also known as Adenosine Triphosphatase functions as an enzyme that catalyzes the hydrolysis of ATP into ADP and inorganic phosphate. This process releases energy which is essential for various cellular activities. ATPases come in various forms including F-type V-type and P-type each with specific roles in cellular energy metabolism. These enzymes are expressed widely in cellular membranes and organelles such as mitochondria chloroplasts and the plasma membrane. The typical molecular mass of ATPase depends on the specific type but it generally ranges from 70 to 100 kDa for individual subunits.
Biological function summary
Enzymes in the ATPase family provide energy required for transport processes signal transduction and muscle contraction. They are essential components of large protein complexes. For example the F1F0 ATP synthase complex in mitochondria and chloroplasts is a pivotal participant in ATP production through oxidative phosphorylation and photophosphorylation. ATPase activity is measured using various assays such as malachite green assay to determine ATPase function by detecting released inorganic phosphate.
Pathways
ATPases are important for energy transduction in key biological pathways like cellular respiration and photosynthesis. They participate in the ATP synthase pathway in mitochondria and chloroplasts and the calcium signaling pathway. These pathways involve interaction with proteins like cytochrome c oxidase in the electron transport chain and calmodulin in calcium homeostasis. ATPase activity assays such as the ATP hydrolysis assay assist researchers in understanding these pathways better by quantifying ATPase activity.
ATPases show associations with conditions like Alzheimer's disease and cystic fibrosis. Abnormalities in ATPase function can disrupt cellular energy balance or ion transport leading to neurological and respiratory pathologies. Mutations in ATP7B a copper-transporting ATPase relate to Wilson's disease leading to improper copper homeostasis. Analyzing ATPase activity and function in disease models helps researchers unveil these molecular mechanisms and potential therapeutic targets.
