Key features and details
- Assay type: Semi-quantitative
- Detection method: Luminescent
- Platform: Microplate reader
- Assay time: 30 min
- Sample type: Adherent cells, Suspension cells, Tissue
- Sensitivity: 100 cells/well
Product nameADP/ATP Ratio Assay Kit (Bioluminescent)
Sample typeTissue, Adherent cells, Suspension cells
Sensitivity< 100 cells/well
Assay time0h 30m
ADP/ATP Ratio Assay Kit (Bioluminescent) ab65313 is based on the bioluminescent detection of ADP and ATP levels. It can be used for a rapid screening of apoptosis, necrosis, growth arrest, and cell proliferation simultaneously in mammalian cells.
In the ADP/ATP assay protocol, luciferase catalyzes the conversion of ATP and luciferin to light, which in turn can be measured using a luminometer or Beta Counter. The ADP level is measured by its conversion to ATP that is subsequently detected using the same reaction.
The assay can be fully automatic for high throughput and is highly sensitive (detects 100 mammalian cells/well).
ADP/ATP Ratio assay protocol summary:
- transfer suspension cells, or nucleotide releasing buffer treated adherent cells, to plate
- add ATP reaction mix and incubate for 2 min
- analyze with luminescence plate reader to measure ATP
- after preparing ADP reaction mix and measuring luminescence levels again, add ADP reaction mix to same wells and incubate for 2 min
- analyze with luminescence plate reader to measure ADP
Changes in the ADP/ATP ratio have been used to differentiate the different modes of cell death and viability. Increased levels of ATP and decreased levels of ADP have been recognized in proliferating cells. In contrast, decreased levels of ATP and increased levels of ADP are recognized in apoptotic cells. The decrease in ATP and increase in ADP are much more pronounced in necrosis than apoptosis.
Review the metabolism assay guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 200 tests ADP Converting Enzyme (Lyophilised) Blue Cap 1 vial ATP Monitoring Enzyme (Lyophilised) 1 vial Enzyme Reconstitution Buffer 1 x 2.15ml Nucleotide Releasing Buffer 1 x 50ml
RelevanceThe changes in ADP/ATP ratio have been used to differentiate the different modes of cell death and viability. Increased levels of ATP and decreased levels of ADP have been recognized in proliferating cells. In contrast, decreased levels of ATP and increased levels of ADP are recognized in apoptotic cells. The decrease in ATP and increase in ADP are much more pronounced in necrosis than apoptosis.
Quantitation of glutamate levels (A) using ab138883 and intracellular ATP (B) using ab65313 in D. hydrothermalis cells grown under different pressure conditions.
ADP/ATP ratio in cystathionine-beta-synthase slienced A2780 (Ovarian cancer cell line) cells and AOAA (aminooxyacetic acid) treated A2780 cells were measured using ADP/ATP ratio assay kit (ab65313).
ADP/ATP ratio were examined in both narciclasine (ncls) and vehicle (veh) treated C2C12 myotubes with or without PA treatment using ADP/ATP ratio assay kit (ab65313).
ab65313 has been referenced in 63 publications.
- Zalewska A et al. The Effect of N-Acetylcysteine on Respiratory Enzymes, ADP/ATP Ratio, Glutathione Metabolism, and Nitrosative Stress in the Salivary Gland Mitochondria of Insulin Resistant Rats. Nutrients 12:N/A (2020). PubMed: 32059375
- Scheuermann U et al. Sirtuin-1 expression and activity is diminished in aged liver grafts. Sci Rep 10:11860 (2020). PubMed: 32681076
- Sun C et al. Re-equilibration of imbalanced NAD metabolism ameliorates the impact of telomere dysfunction. EMBO J 39:e103420 (2020). PubMed: 32935380
- Matsuura S et al. Platelet Dysfunction and Thrombosis in JAK2V617F-Mutated Primary Myelofibrotic Mice. Arterioscler Thromb Vasc Biol 40:e262-e272 (2020). PubMed: 32814440
- Lieber T et al. Mitochondrial fragmentation drives selective removal of deleterious mtDNA in the germline. Nature 570:380-384 (2019). PubMed: 31092924