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) is designed to be a robust, simple method which utilizes 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.


    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.


    Visit our FAQs page for tips and troubleshooting.


    Review our 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.

  • Notes

    ATP is the primary energy currency of living systems. Virtually all energy requiring processes utilize the chemical energy stored in the phosphate bond of ATP. ATP is formed exclusively in the mitochondria and a variety of genetic diseases can affect ATP formation in the mitochondria.

     

Properties

Associated products

Images

  • Functional studies - ab83355
    Functional studies - 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).

  • Functional studies - ab83355
    Functional studies - 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).

  • Functional Studies - ATP Colorimetric/Fluorometric Assay Kit (ab83355)
    Functional Studies - ATP Colorimetric/Fluorometric Assay Kit (ab83355)
    Example of fluorometric ATP assay standard curve.
  • Functional Studies - ATP Colorimetric/Fluorometric Assay Kit (ab83355)
    Functional Studies - ATP Colorimetric/Fluorometric Assay Kit (ab83355)
    Example of colorimetric ATP assay standard curve.
  • Functional Studies - ATP Colorimetric/Fluorometric Assay Kit (ab83355)
    Functional Studies - ATP Colorimetric/Fluorometric Assay Kit (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 47 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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34 reviews or Q&A

10
Votes: Highest First

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

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

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

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.

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?

This kit will detects total ATP.

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?

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 de...

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

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

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 ...

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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...

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ATP measurement after siRNA treatment in cancer cells

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
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.
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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
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.
abreview image
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Mrs. Fotini MOUTH

Verified customer

Submitted Nov 23 2017

We want to examine the ATP levels in cell cultures after stimulating the cells in highly oxygenated cell media. Can high oxygen levels affect the results when using the ATP assay?

This assay utilizes an oxired probe reaction. ATP is used during enzymatic phosphorylation of glycerol and the byproduct of this reaction eventually oxidizes the probe to form Resorufin which is measured by absorbance or fluorescence. The higher the AT...

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I have two questions about sample preparation/stability. I will be using harvested skeletal muscles for this assay.

1) The instruction booklet says to homogenize the sample with ATP assay buffer first and then deproteinize using PCA and neu...

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Directly homogenizing in PCA makes sure that all proteins are precipitated and enzymes are deactivated right away. This is a good option for muscles since they are highly metabolically active tissues. This will also be helpful for you since this would ...

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