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    products/assay-kits/adpatp-ratio-assay-kit-bioluminescent-ab65313.pdf

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ADP/ATP Ratio Assay Kit (Bioluminescent) (ab65313)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (2)Q&A (39)References (108)

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Functional studies - ab65313
  • Functional studies - ab65313
  • Functional studies - ab65313

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

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Overview

  • Product name

    ADP/ATP Ratio Assay Kit (Bioluminescent)
  • Detection method

    Luminescent
  • 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 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

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K255 ADP/ATP Ratio Bioluminescence Assay Kit, ApoSENSOR. K255-200 is the same size as the 200 test size of ab65313.

    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.

    Related assays

    Review the cell health assay guide to learn about kits to perform a cell viability assay, cytotoxicity assay and cell proliferation assay. 

    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.

  • Platform

    Microplate reader

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

    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Other Metabolism Assay
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Intermediary Metabolism Kits
  • 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.

Associated products

  • Assay kits

    • Luminescent ATP Detection Assay Kit (ab113849)
    • ATP Assay Kit (Colorimetric/Fluorometric) (ab83355)

Images

  • Functional studies - ab65313
    Functional studies - ab65313Image from Amrani A et al., PLoS One 9(9). Fig 5A. doi: 10.1371/journal.pone.0106831 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Quantitation of glutamate levels (A) using ab138883 and intracellular ATP (B) using ab65313 in D. hydrothermalis cells grown under different pressure conditions.

  • Functional studies - ab65313
    Functional studies - ab65313Bhattacharyya S.,PLoS One 8(11), Fig 5d & f. doi: 10.1371/journal.pone.0079167 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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).

     

  • Functional studies - ab65313
    Functional studies - ab65313Image from Julien S.G., PLoS Biol 15(2), Fig 8C. doi: 10.1371/journal.pbio.1002597. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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). 

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (108)

Publishing research using ab65313? Please let us know so that we can cite the reference in this datasheet.

ab65313 has been referenced in 108 publications.

  • You B  et al. AMPK-mTOR-Mediated Activation of Autophagy Promotes Formation of Dormant Polyploid Giant Cancer Cells. Cancer Res 82:846-858 (2022). PubMed: 34965934
  • Yim GW  et al. Long Non-coding RNA LOC285194 Promotes Epithelial Ovarian Cancer Progression via the Apoptosis Signaling Pathway. In Vivo 36:121-131 (2022). PubMed: 34972707
  • Wang SW  et al. Limonin, an AMPK Activator, Inhibits Hepatic Lipid Accumulation in High Fat Diet Fed Mice. Front Pharmacol 13:833705 (2022). PubMed: 35140621
  • Lv J  et al. Mitochondrial homeostasis regulates definitive endoderm differentiation of human pluripotent stem cells. Cell Death Discov 8:69 (2022). PubMed: 35177589
  • Sosnowski DK  et al. Changes in the Left Ventricular Eicosanoid Profile in Human Dilated Cardiomyopathy. Front Cardiovasc Med 9:879209 (2022). PubMed: 35665247
View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

ADP/ATP Ratio Assay in M. tuberculosis

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
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.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Gregory Olson

Verified customer

Submitted Oct 06 2017

Very sensitive and easy to use

Excellent Excellent 5/5 (Ease of Use)
Abreviews
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.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Craig Beall

Verified customer

Submitted Sep 17 2014

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