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Cell Viability Assay Kit (Bioluminescent) (ab65314)

Submit a review Q&A (8)References (11)

Key features and details

  • Assay type: Direct
  • Detection method: Luminescent
  • Platform: Microplate reader
  • Assay time: 30 min
  • Sample type: Adherent cells, Suspension cells
  • Sensitivity: 10 cells/well

Overview

  • Product name

    Cell Viability Assay Kit (Bioluminescent)
    See all Cell Viability kits

  • Detection method

    Luminescent

  • Sample type

    Adherent cells, Suspension cells

  • Assay type

    Direct

  • Sensitivity

    > 10 cells/well

  • Range

    10 cells/well - 100 cells/well

  • Assay time

    0h 30m

  • Product overview

    Abcam's Cell Viability Assay Kit (Bioluminescent) utilizes bioluminescent detection of the ATP levels for a rapid screening of apoptosis and cell proliferation simultaneously in mammalian cells. The assay utilizes luciferase to catalyze the formation of light from ATP and luciferin, and the light can be measured using a luminometer or Beta Counter. The assay can be fully automatic for high throughput (10 seconds/sample). The high sensitivity of this assay has led to many other applications for detecting ATP production in various enzymatic reactions, as well as for detecting low level bacterial contamination in samples such as blood, milk, urine, soil, and sludge.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K254 ApoSENSOR™ ATP Cell Viability Bioluminescence Assay Kit. K254-200 is the same size as the 200 test size of ab65314.

    Cell death (especially apoptosis) is an energy-dependent process that requires ATP. As ATP levels fall to a point where the cell can no longer perform basic metabolic functions, the cell will die. A typical apoptotic cell exhibits a significant decrease in ATP level. Therefore, loss of ATP level in cell has been used as an indicator of cell death. In contrast, cell proliferation has been recognized by increased levels of ATP.

  • Platform

    Microplate reader

Properties

References (11)

Publishing research using ab65314? Please let us know so that we can cite the reference in this datasheet.

ab65314 has been referenced in 11 publications.

  • Poprawa I  et al. Ovaries and testes of Lithobius forficatus (Myriapoda, Chilopoda) react differently to the presence of cadmium in the environment. Sci Rep 12:6705 (2022). PubMed: 35469038
  • Qing Y  et al. R-2-hydroxyglutarate attenuates aerobic glycolysis in leukemia by targeting the FTO/m6A/PFKP/LDHB axis. Mol Cell 81:922-939.e9 (2021). PubMed: 33434505
  • Veremeyko T  et al. Neuronal extracellular microRNAs miR-124 and miR-9 mediate cell-cell communication between neurons and microglia. J Neurosci Res 97:162-184 (2019). PubMed: 30367726
  • Chen F  et al. YY1 regulates skeletal muscle regeneration through controlling metabolic reprogramming of satellite cells. EMBO J 38:N/A (2019). PubMed: 30979776
  • Rekha B  et al. Chronic intake of 4-Methylimidazole induces Hyperinsulinemia and Hypoglycaemia via Pancreatic Beta Cell Hyperplasia and Glucose Dyshomeostasis. Sci Rep 8:17037 (2018). PubMed: 30451881
View all Publications for this product

Customer reviews and Q&As

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1-8 of 8 Abreviews or Q&A

Question

I want to test for viability in cell culture supernatants. We are using RPMI + 10% FBS in our assay. Will this have an effect in the readout?

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Answer



I've confirmed with the lab, and you can use this kind of media but be sure to use a control media with the same amount of RPMI and FBS to account for any signals generated due to these 2 in your sample readings. Just subtract the control readings from the other readings.

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Question

I want to test for viability in cell culture supernatants. We are using RPMI + 10% FBS in our assay. Will this have an effect in the readout?



abcam.com

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Answer

Hello!

You can use a control media with the same amount of RPMI and FBS to account for any signals generated due to these 2 in your sample readings. Just subtract the readings you get in the controls from the standards.

Please let me know if you have any further questions.

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Question

I see that the readout would be ATP in the media; this means that non-viable (or dead) cells would have lower ATP readings in the media compared to live cells.

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Answer


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

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Question

Can you please suggest a product for which I can test cell viability using cell supernatants?

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Answer

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

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Question


I have one question regarding the cell viability assay kit (ab65314).
In the manual is not very clear to me if each sample should be read for 1 minute or not. I found 2 different indications:

The assay can be fully automatic for high throughput (10 seconds/sample) and is extremely sensitive (detects
10-100 mammalian cells/well) (page 3)
Read the sample for 1 minute in a luminometer (page 7).

And another question: if each sample such be read for 1 minute and I have a full plate it will take more than 1 hour to read the whole plate. Is the signal stabile enough?

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Answer

Thank you for contacting us and for asking your question about the Cell Viability Assay Kit instructions.

The instruction on page 7 of the protocol to read the samples for one minute after adding the enzyme is mistaken. It should read:

"3. Add 5 μL ATP Monitoring Enzyme into the cell lysate. After 1 minute, read the samples in a luminometer."

The example given earlier in the protocol for high-throughput reading at 10 seconds per sample is correct but is only a suggestion. The amount of time required for reading each sample will vary depending on the instrument.

I apologize for the confusion. Please let me know if you have any questions.

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Question

Sure you suggest a luminometer but a beta counter as well in the protocol, for this reason I wrote you asking for more information about the Beta counter setup.

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Answer

Thank you for your reply. I can confirm that conventional scintillation counters can not be used for measurements with this kit. However, when Beta Counter is used it should be programmed in the “out of coincidence” (or Luminescence mode) for measurement. So if they have this setting where they can program this, then the beta counter may be used. I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Question

Dear ABcam, concerning the protocol, I'd like to know if I can use a Beckman Coulter LS6500 Multipurpose Scintillation Counter to carry out my experiment and how i could setup my instrument. Thank you

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Answer

Thank you for your enquiry. I can confirm that the read out of ab65314  Cell Viability Assay Kit is via luminescence. Therefore, we would suggest to use the indicated equipment which is a luminometer.  Theoretically a luminometer and a scintillation counter are different. I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Question

I have a question concerning the ATP assay kit (ab65314. I work with isolated mitochondria (from mammalian tissue) and my question is whether your kit will work in my case.

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Answer

Thank you for your enquiry. I have contacted the originator of this kit who have confirmed that it can be used for measuring the ATP in mitochondrial lysates. Once you have produced your mitoochondria lysates, you can follow the kit protocol and get the ATP value. The kit protocol can be found by clicking on the protocols tab at the top of the online datasheet. The kit procedure will be the top link on the list of protocols provided. I hope this is helpful to you. Should you have any further questions, please do not hesitate to contact us.

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