Overview

  • Product name
    ADP/ATP Ratio Assay Kit (Bioluminescent)
  • Sample type
    Tissue, Adherent cells, Suspension cells
  • Assay type
    Semi-quantitative
  • Sensitivity
    < 100 cells/well
  • Assay time
    0h 30m
  • Product overview

    ADP/ATP Ratio Assay Kit (Bioluminescent) (ab65313) is based on the bioluminescent detection of the ADP and ATP levels in the sample of interest for a rapid screening of apoptosis, necrosis, growth arrest, and cell proliferation simultaneously in mammalian cells. In this assay, luciferase catalyzes the conversion of ATP and luciferin to light, which in turn can be measured using a luminometer or Beta Counter. 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).

     

    Visit our FAQs page for tips and troubleshooting.

  • Notes

    The 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.

  • Tested applications
    Suitable for: Functional Studiesmore details

Properties

  • Storage instructions
    Store 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
  • Research areas
  • Relevance
    The 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.

Applications

Our Abpromise guarantee covers the use of ab65313 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies Use at an assay dependent dilution.

Protocols

References

This product has been referenced in:
  • Julien SG  et al. Narciclasine attenuates diet-induced obesity by promoting oxidative metabolism in skeletal muscle. PLoS Biol 15:e1002597 (2017). Read more (PubMed: 28207742) »
  • Xue M  et al. Supramolecular Probes for Assessing Glutamine Uptake Enable Semi-Quantitative Metabolic Models in Single Cells. J Am Chem Soc 138:3085-93 (2016). Read more (PubMed: 26916347) »

See all 22 Publications for this product

Customer reviews and Q&As

ADP/ATP Ratio Assay in M. tuberculosis

Excellent Excellent 5/5 (Ease of Use)
Abreviews
We tested ab65313 ADP/ATP Ratio Assay in M. tuberculosis (Mtb) strains H37Rv and 18b. The main goal was to determine if the Nucelotide Releasing Buffer (NRB) was able to cause release of nucleotides from Mtb with or without additional disruption (bead beating).
Experimental set-up:
Cells: Mtb strains H37Rv and 18b were grown in 7H9 media and harvested at an OD600 of ~0.4. 175 Million total bacteria were used from each strain.
Control: 7H9 alone
Treatments:
- "Lysis": NRB (5-10 minutes)
- "Beads": NRB (5-10 minutes) + bead-beading (speed 6.60 - 3 x 30s runs)
Measurement:
ab65313 on a Pherastar Plus plate reader on bioluminescent setting
Standard Curve:
ATP disodium salt (ab120385) dilutions to create a standard curve.
Results:
Standard curve: log10(RLU) = 3.739 + 0.938 * log10(ATPconc)
Boxplot of ADP and ATP concentrations on two technical replicates on each of two biological replicates. #N.B. The gain setting on the luminometer resulted in ADP readings (Data D of the kit) for the bead-beating groups that were off-scale and thus non-quantifiable, but still plotted in the attached figure.
Discussion:
NRB alone did not lead to appreciable release of nucleotides from two strains of Mtb. The addition of bead beating to NRB led to significant increases in measured concentrations of both nucleotides in both strains.
Username

Mr. Gregory Olson

Verified customer

Submitted Oct 06 2017

The assay buffer has high concentration of detergent-hence Bradford assay is not recommended. We advise using the BCA assay for protein measurement when using this kit.
This assay is performed on cell lysates, so it would be difficult to label the ...

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Thank you for contacting us. Regarding your questions from yesterday:

1. Why is data C required after data B acquisition?

The two readings should be similar and perhaps data C is not necessary but the ATP signal may go down slightly w...

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Very sensitive and easy to use

Excellent Excellent 5/5 (Ease of Use)
Abreviews
We have used this product extensively to measure changes in the ATP/ADP ratio in response to changes in glucose availability and during stress tests (H2O2 treatment). We have some recommendations below for use:

1) Use a black-walled 96-well plate to reduce plate phosphorescence
2) If possible, use ~1,000 cells per well. If you can do a seeding density curve, you will notice that the ATP/ADP ratio drops with increasing cell number. This is due to glucose depletion for the media. Less cells will give you a better charged cell.
3) Following on from point 2, change the media 1-2 hours before starting your experiment. This will replenish the glucose in the media. We found this to be very important when trying to utilise physiological glucose concentrations (2.5-10 mM glucose).

Reliable kit and easy to use.
Username

Dr. Craig Beall

Verified customer

Submitted Sep 17 2014

The signal should be fairly stable as long as the sample is protected from light. But we suggest measuring after 2 mins to make sure that no light is quenched. Waiting for 10 mins is not advisable since some light can be quenched during this time. For ...

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We can recommend two protocols:
1. Prepare a single cell suspension from the tissues of interest with any method desired (cells have to stay intact). Then treat the cells as instructed in the protocols section for adherent cells: Remove cultur...

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Please see the following publication which may be of use to you:

Diguet N et al. Muscle creatine kinase deficiency triggers both actin depolymerization and desmin disorganization by advanced glycation end products in dilated cardiomy...

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Thank you for your inquiry.

I am happy to confirm that ab65313 can be used with mouse liver tissue samples.

This kit can be used to measure the ADP/ATP Ratio, AMP is not measured.

If you are planning to use this kit with t...

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Yes, you are correct. Lower ATP readings would indicate a decrease in cell viability.

Ab65314 is recommended for cell viability. For media, use the same protocol as mentioned for the suspension cells in step 2.a.

1-10 of 41 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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