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Luminescent ATP Detection Assay试剂盒(ab113849)

Reviews (7)Q&A (30)References (234)
Functional Studies - Luminescent ATP Detection Assay Kit (ab113849)
  • Example of ATP standard curve using an opaque white plate
  • ATP Luminescence Assay using ab113849
  • Simultaneous quantification of mitochondrial respiration and glycolytic flux
  • Extracellular detection of ATP (ab113849)

Key features and details

  • Assay type: Quantitative
  • Detection method: Luminescent
  • Platform: Microplate reader
  • Assay time: 30 min
  • Sample type: Adherent cells, Suspension cells

概述

  • 产品名称

    Luminescent ATP Detection Assay试剂盒
    参阅全部 ATP 试剂盒

  • 检测方法

    Luminescent

  • 样品类型

    Adherent cells, Suspension cells

  • 检测类型

    Quantitative

  • 检测时间

    0h 30m

  • 产品概述

    Luminescent ATP Detection Assay Kit (ab113849) is used to measure the level of ATP within the cell. The luminescent ATP assay protocol involves lysis of the cell sample, addition of luciferase enzyme and luciferin, and measurement of the emitted light using a tube or microplate-based luminometer.


    This kit irreversibly inactivates ATP degrading enzymes (ATPases) during the lysis step, ensuring that the luminescent signal obtained truly corresponds to the endogenous levels of ATP.


    Luminescent ATP assay protocol summary:
    - add ATP standard into standard wells and media into control wells in same plate containing cells to be analyzed
    - add detergent solution and incubate for 5 min to lyse cells and stabilize ATP
    - add substrate solution and incubate for 5 min
    - store plate in dark for 10 min
    - analyze on luminescence plate reader


    Special Handling Instructions for the ATP Detection Assay Kit


    ATP can be found in cells and microbiota on many surfaces. To prevent unintended background, it is recommended to clean bench surfaces and all pipettes to be used during the experiment with 10% bleach. Use of gloves first cleaned by either using 70% ethanol or by changing them frequently is recommended. Use tips and containers that are clean and sterile, such as ATP and nuclease-free consumables.  Do not leave reagents or the plate opened while working or during assay incubation

  • 说明

    Total levels of cellular ATP can be used to assess cell viability, cell proliferation and cytotoxicity of a wide range of compounds and biological response modifiers.

    We also offer a very popular alternative colorimetric/fluorometric ATP assay kit ab83355 based on the phosphorylation of glycerol.

    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.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • 平台

    Microplate reader

性能

图片

  • Functional Studies - Luminescent ATP Detection Assay Kit (ab113849)
    Functional Studies - Luminescent ATP Detection Assay Kit (ab113849)D?ini?, Tamara and Norbert A Dencher., Oxidative medicine and cellular longevity?vol. 2018 7567959., Fig 6, doi:10.1155/2018/7567959

    Total cellular ATP concentration. ATP in SH-SY5Y cells cultivated at 21% and 5% O2 24 h after treatment with A? peptide and/or 18 h X-ray irradiation, normalized to cell count, and compared to respective controls. ATP concentration was about 1.3- to 1.8-fold higher at all conditions in cells cultivated at 5% O2 compared to 21% O2. Combination of A? peptide treatment and irradiation resulted in a significantly increased (~1.5-fold) ATP concentration at 5% O2 compared to the control. Samples were measured at least in duplicates (n = 2-4) in three independent experiments (N = 3). Mean ± SEM analyzed by two-way ANOVA with Tukey's multiple comparison test with p < 0 05 considered as significant. (∗∗p < 0 01).

  • Example of ATP standard curve using an opaque white plate
    Example of ATP standard curve using an opaque white plate

    The ATP standard curve was prepared as described in the protocol. Background-subtracted data values (mean +/- SD) are graphed.

  • ATP Luminescence Assay using ab113849
    ATP Luminescence Assay using ab113849
    ab113849 ATP detection kit cytotoxicity data. 25000 HepG2 cells were seeded into each well, allowed to adhere and treated for 4 hours with 25µM rotenone and vehicle control (DMSO) in glucose based complete media. After treatment, cells were lysed, exposed to the ATP substrate solution and signal was measured on a luminescent counter. Mean and standard deviation is plotted for 3 replicates from each condition. Rotenone induces cytotoxicity in HepG2 cells.
  • Simultaneous quantification of mitochondrial respiration and glycolytic flux
    Simultaneous quantification of mitochondrial respiration and glycolytic flux

    Cellular Energy Flux for HepG2 cells (seeded at 65,000 per well), treated with a combination of drug compounds modulating the ETC (Antimycin A [1 µM] and FCCP [2.5 µM]), shown as a percentage relative to untreated control cells. Comparative measurements were taken with Extracellular Oxygen Consumption Assay (ab197243) (white column) and Glycolysis Assay [Extracellular acidification] (ab197244) (black column) show the shift between mitochondrial respiration and glycolysis and the cellular control of energy (ATP; measured 1h post-treatment using Luminescent ATP Detection Assay kit (ab113849) (striped column)).

  • Extracellular detection of ATP (ab113849)
    Extracellular detection of ATP (ab113849)This image is courtesy of an Abreview submitted by Heiko Lemcke.

    Analysis of the release of ATP by connexin hemichannels in stem cells using ATP luminescence kit (ab113849).

    Cells were cultured in HBSS to induce hemichannel opening. Calcium and GAP-inhibitor were used to trigger hemichannel closure.
    After two hours the supernatant was collected and ATP was measured according to the protocol (detergent was also applied).

    Calcium treatment and inhibition by GAP decreased ATP concentration, compared to HBSS control. Graph shows data of three independent experiments.

文献 (234)

发表研究结果有使用 ab113849？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab113849 被引用在 234 文献中.

  • Sittewelle M & Royle SJ Passive diffusion accounts for the majority of intracellular nanovesicle transport. Life Sci Alliance 7:N/A (2024). PubMed: 37857498
  • Najjar RS  et al. Raspberry polyphenols target molecular pathways of heart failure. J Nutr Biochem 124:109535 (2024). PubMed: 37984734
  • Liu J  et al. Icariin ameliorates glycolytic dysfunction in Alzheimer's disease models by activating the Wnt/β-catenin signaling pathway. FEBS J 291:2221-2241 (2024). PubMed: 38400523
  • Kersten K  et al. Uptake of tumor-derived microparticles induces metabolic reprogramming of macrophages in the early metastatic lung. Cell Rep 42:112582 (2023). PubMed: 37261951
  • Boyd RA  et al. The heat shock protein Hsp27 controls mitochondrial function by modulating ceramide generation. Cell Rep 42:113081 (2023). PubMed: 37689067
View all Publications for this product

客户评价及客户问答

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1-10 of 37 Abreviews or Q&A

ATP measurement in cell culture supernatant

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
BV-2 microglial cells were plated in 12-well plates and at the end of the experiment the culture medium was collected and centrifuged. ATP was quantified in supernatant in control and Yoda-1 incubated cells, following the protocol provided in the datasheet.
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.

Abcam user community

Verified customer

提交于 Apr 11 2024

Luminescent ATP Detection in tissue

 Good Good 4/5 (Ease of Use)
Abreviews
abreview image
We assessed ATP differences in the hearts of animals that underwent either left anterior descending coronary artery ligation to induce heart failure (HF), or a sham procedure as a control. The ATP assay worked well with a modified protocol (detailed below). As expected, HF animals had substantially lower ATP content compared with sham animals (~42% reduction).

Protocol modifications: 5 mg of tissue (left ventricle in our case) was homogenized in a glass dounce homogenizer with 500 uL of detergent on ice. No inhibitors were added. After samples were homogenized, they were allowed to sit at room temperature for 10 min. Samples were then centrifuged for 5 min at 13,000 g at 4 degrees C. Supernatant was placed into fresh tubes. Protein of supernatant was quantified. A white, flat bottom 96-well plate (bottom was white, not clear) was utilized. Samples were added to wells at a concentration of 10 ug/mL, and final volume in the well was 50 uL as per the Abcam protocol. Following this, the procedures were identical to the Abcam protocol.
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.

Rami Najjar

Verified customer

提交于 Jul 20 2022

Cellular ATP measurement

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
This kit is very simple and easy to use.
Always had consistent results.
I highly recommend this kit.
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.

Abcam user community

Verified customer

提交于 Sep 11 2020

Changes in ATP levels in BV2 microglia

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
The kit was used to analyze changes in ATP levels in BV2 microglia cells. The cells were treated with 1µM lysophosphatidic acid (LPA) for the indicated time points. Rapid increase in the levels of ATP were observed at short time points. The kit was easy to use and get the results in less than an hour. We observed some variation between triplicates but did not affect much the outcome.
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.

Abcam user community

Verified customer

提交于 Jun 14 2019

ATP levels in microglia cells

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
The kit is very easy to use and the results are reproducible. We used it to evaluate changes in ATP levels in primary murine microglia (in BV2 cell line as well) after treatment with various concentrations of lysophosphatidic acid (LPA).
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. Joanna Plastira

Verified customer

提交于 Oct 11 2018

Extracellular detection of ATP (ab113849)

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
We have analyzed the release of ATP by connexin hemichannels in stem cells using the ATP luminescence kit (ab113849).

Cells were cultured in HBSS to induce hemichannel opening. Calcium and GAP-inhibitor were used to trigger hemichannel closure.
After two hours the supernatant was collected and ATP was measured according to the protocol (we also applied the detergent).

As expected calcium treatment and inhibition by GAP decreased ATP concentration, compared to HBSS control. Graph shows data of three independent experiments.

In summary, we can recommend the ATP luminescence kit (ab113849) for measuring extracellular ATP concentrations.
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.

Heiko Lemcke

Verified customer

提交于 Jul 31 2017

Measurement of extracellular ATP levels

 Good Excellent 5/5 (Ease of Use)
Abreviews
abreview image
I enrolled to an Abcam trial to check the suitability of this ATP luminescence kit (ab113849) to detect extracellular ATP levels in cancer cells.

I personally recommend this kit to measure extracellular ATP levels for its ease of use and for the kindly attention and help I received from the Abcam scientific support specialists. When measuring extracellular ATP levels, it is important to avoid light exposure as much as possible and measure the supernatant without FBS and without phenol-red.

Brief description of the protocol:
I seeded 50.000 murine breast-to-brain cancer cells in duplicates or triplicates in 96-well-plate. Once they are attached I starve them in FBS-free and phenol red-free media.

24 hours after starving I collected the supernatant (100 ul) and transfer it to a white opaque plate. There I followed the protocol proposed in the booklet with only one difference: I do not use detergent.

Using the same conditions (cell line, passage, starving time and machine) I repeated 3 times with the same cancer cell line and the results I got are 12,89 nM ATP, 12,30 nM ATP and 7,3 nM ATP.

Regarding the conditions I use, I repeated this procedure several times and I chose this condition (24-hours starvation in phenol red-free and serum-free media) because if I starve them for less hours the luminescence units I get from my samples are super low. If I starve them with HBBS overnight the cells look ugly the following day. Moreover I also tried to add detergent in the media but then the RLU I got were more variable in the same biological replicate. There is also higher variability of RLU if I use media with FBS. I did not try to use media with phenol red because I know it interferes with the luminescence readout. To sum up, I got less variability when I starve them for 24 hours with DMEM without FBS and without phenol red.
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. Julian Anthes

Verified customer

提交于 Jul 07 2017

Question

ab113849
what type of 96 well plates are required. Would clear ones be ok?

Read More
Answer

I have looked into this further and can confirm that in testing clear plates have been used with no major issues. However if you do have black plates, that may be more optimal.

Read More

Question

Inquiry: Hello, I've used the Luminescent ATP Detection Assay Kit to determine the concentration of ATP in E. coli cells. The kit worked perfectly. My question is, will the lysis protocol denature ATP binding proteins such that they release ATP allowing it to take part in the luminescence reaction? Has any work been conducted on this?

Read More
Answer

We haven’t addressed this question experimentally with the reagents.  The answer is probably.  The lysis reagent is denaturing so bound ATP may be made available to the luciferase. This would be difficult to be definitive and different ATP binding proteins may behave differently.

Read More

Question

Is the Luminescent ATP Detection Assay Kit (ab113849) suitable for use with plasma samples, or is it purely for use with cell samples?

Read More
Answer


In theory it should work, but we don’t know for sure as we have not tested it. Therefore, if you are willing to test and publish the data in the form of review on our website then I would like to invite you to our Abtrial programme.

Read More

1-10 of 37 Abreviews or Q&A

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