ATP Assay Kit (Colorimetric/Fluorometric) (ab83355)

Reviews (4)Q&A (32)References (55)

Overview

  • Product name
    ATP Assay Kit (Colorimetric/Fluorometric)
    See all ATP kits
  • Detection method
    Colorimetric/Fluorometric
  • Sample type
    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate
  • Assay type
    Quantitative
  • Sensitivity
    < 1 µM
  • Assay time
    1h 00m
  • Product overview

    ATP Assay Kit (Colorimetric/Fluorometric) (ab83355) use a robust, simple method; the ATP assay protocol relies on the phosphorylation of glycerol to generate a product that is easily quantified by colorimetric (ODmax = 570 nm) or fluorometric (Ex/Em = 535/587 nm) methods.


    This kit can detect as low as 1 µM of ATP in various samples.


    ATP assay protocol summary:
    - add samples (deproteinized) and standards to wells
    - add reaction mix and incubate for 30 min at room temp
    - analyze with microplate reader


    If you require a more sensitive product, we recommend Luminescent ATP Detection Assay Kit (ab113849), which can detect as low as 1 pM of ATP.

  • Notes

    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

Associated products

Images

  • ATP assay performed with ab83355
    ATP assay performed with ab83355Sanokawa-Akakura R et al., PLoS One 9(9), fig4b. doi: 10.1371/journal.pone.0108537

    The chart shows a comparison of ATP levels of HepG2 treated with 0, 10 and 100 µM for 48 hours, DRH2O2 W1 (damage recovered cells using hydrogen peroxide with a recovery time of one week) HepG2 cells and MDA-MB-231 cells treated with 0, 10 and 100 µM of NaHS for 48 hours. Data is shown as percent of ATP levels in untreated cells. ATP levels were determined using ATP assay kit (ab83355).

  • ATP assay performed with ab83355
    ATP assay performed with ab83355Image from Shi Z et al., PLoS One 9(8), Fig 2 doi: 10.1371/journal.pone.0105675.

    MCF-7 cells are transfected with vector, osteopontin-a, osteopontin-c or osteopontin -a plus -c. Cells are plated on poly-HEMA and seeded at 4 x 105 cells per well and incubated for two days under standard culture conditions. ATP levels are measured using ATP assay kit (ab83355).

  • ATP assay performed with ab83355
    ATP assay performed with ab83355
    Example of fluorometric ATP assay standard curve.
  • ATP assay performed with ab83355
    ATP assay performed with ab83355
    Example of colorimetric ATP assay standard curve.
  • ATP assay performed with ab83355
    ATP assay performed with ab83355

    Quantitation of ATP in fish liver (2.5µl of 10 times diluted sample), fish muscle (5µl of 10 times diluted sample) and Jurkat cell lysate (5 ul) using fluorometric assay. Samples were spiked with known amounts of ATP (300pmol).

Protocols

References

This product has been referenced in:
  • Han SJ  et al. IDH2 deficiency increases the liver susceptibility to ischemia-reperfusion injury via increased mitochondrial oxidative injury. Redox Biol 14:142-153 (2018). Read more (PubMed: 28938192) »
  • Zou X  et al. Renal scattered tubular-like cells confer protective effects in the stenotic murine kidney mediated by release of extracellular vesicles. Sci Rep 8:1263 (2018). Read more (PubMed: 29352176) »
See all 55 Publications for this product

Publishing research using ab83355? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 36 Abreviews or Q&A

Question

Can you please let me know if desferrioxamine interferences with the ATP assay?

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Answer

Unfortunately, we have not tested the interference of desferrioxamine with this kit.

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Question

Do you have any recommendations for the type of 96 well plate to use for the fluorometric assay? Black, flat bottomed, low/high binding?

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Answer

For the colorimetric assay, use a clear plate. For the fluorometric assay a clear or a white plate can be used. Use flat bottom plates.

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Question

I want to use ATP Assay Kit to determine the ATP level in bone marrow cells. My question is the ATP level measured by this kit is total ATP (cellular plus inside mitochondria) or only cellular ATP?

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Answer

This kit will detects total ATP.

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Question

The kit instructions are pretty vague on the cell lysis step.  Can you advise us on a methodology for lysing tissue in the ATP Assay Buffer? Specifically for how long, and at what temperature?

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Answer

There are 2 ways you can efficiently lyse your cells for this assay kit. You can either homogenize the 1 x 10^6 cells with 100 ul of the assay buffer using 30-50 passes on ice, check for the homogenization and then deproteinate your samples with the deproteinization kit. Alternatively, you can just lyse your cells in the PCA using the protocol mentioned for the deproteinization kit, neutralize the lysate and use it in the wells. In this case you will have to use the assay buffer for diluting the samples in the wells.

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Question

Should protease inhibitors be added to step 1 of the sample prep (lysing cells in ATP assay buffer)?

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Answer

Yes, protease inhibitors can be added during step 1 of the sample preparation step.

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Question

Your protocol mentions that the ATP standard vial is 1 micromol; how can you can make a 10 mMOL stock from 1 Micromol? As you mentioned in page 7 1/100 dilution of 1m MOl (1000 Micro molar or 1000000 nano molar) generates 0,2,4,6,8,10 micro molar not nano molar?

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Answer

The calculations are as follows;

1umol is not micro mole means, it is not molarity it is absolute weight.

1micromol is dilutes in 100ul of water which will give you 1micromol/100ul or 1/100 micromol per ul

Now 10ul of this is diluted further 10 time i.e. 1/100x1/10 = 1/1000 micromol/ul which means 1nanomol/microlitre

1ul = 1 nanomol

2ul = 2 nano mol

10ul = 10 nanomol

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CYP-450 Epoxygenase Derived Epoxyeicosatrienoic Acid Contribute To Reversal of Heart Failure in Obesity-Induced Diabetic Cardiomyopathy via PGC-1 α Activation

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
I tried many different products to get better ATP measurements in Heart tissue of mice but failed then I bought abcam ATP Assay Kit (Colorimetric/Fluorometric) (ab83355) to measure ATP content in heart tissue of mcie which was knockout to PGC1 alpha, as recommended in their datasheets and I never had any problem with it. I got very important data which I used for my research article. I think its great kit to use and have everything in it. I think this is best product to detect ATP content.

Abcam user community

Verified customer

Submitted Aug 07 2018

Testing ab83355 kit in cell culture media

 Good Excellent 5/5 (Ease of Use)
Abreviews
abreview image
We are interested in cuantifying the amount of ATP released to the culture media from primary astrocytes and microglia BV2 after their treatment with different compounds. To that end, we collected the media (DMEM with 1% of penicilin/streptomicin and 4mM of L-glutamine, but without phenol red) and froze them in liquid nitrogen. When we proceeded with the quantification, we thawed the samples on ice and centrifuged them at 13,000 xg for 10 minutes, without applying any deproteinization step. After this, we went on with the assay. We tested the two methodologies available: colorimetric and fluorimetric. Using cells as positive control, we obtained signal by both procedures, but fluorimetric's was the most sensitive, with the hightest signal/background ratio.

Abcam user community

Verified customer

Submitted Jul 31 2018

ATP measurement after siRNA treatment in cancer cells

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
We knockdown a protein which we are studying to effect the cell proliferation.
siRNA knockdown showed significant reduction in ATP generation in cancer cell lines.

Abcam user community

Verified customer

Submitted Mar 27 2018

Test of ab83355 on pancreatic tissue that have undergo cold ischemia

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
We are actually working on the preservation of rat pancreas from ischemia-reperfusion injury that occurs during the retrieval of the organ before a transplantation to a recipient. We are looking for strong ischemia markers and we know from literature that ATP level in pancreatic islet before is a good marker to predict transplantation outcomes (Jae Hyeon K et al., 2009. Transplantation).
We extracted ATP from fresh pancreas that have undergo different time of cold ischemia : 0, 2, 4, 6, 8 and 10h. We also extracted ATP in situ. ATP were extracted in Perchloric acid (PCA-2M) and grind using a Polytron. PCA were removed using potassium hydroxide (KOH – 2M) and pH was adjust around 7-8. Samples were conserved at -80°C before utilization.

mean ATP (pmol/well/mg of tissue) SEM
0H ischemia 424,6105618 90,37421351
2H ischemia 41,0225542 7,565889256
4H ischemia 42,510 5,383
6H ischemia 33,643 2,403
8H ischemia 26,986 2,403
10H ischemia 22,424 1,784
12h ischemia 18,24979888 4,449727198

We obtained concording results with literature : a decrease of ATP level in pancreas with time of cold ischemia. This assay is working on pancreatic rat tissue.

Mrs. Fotini MOUTH

Verified customer

Submitted Nov 23 2017

1-10 of 36 Abreviews or Q&A

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