製品の概要

  • 製品名

    ATP Assay Kit (Colorimetric/Fluorometric)
    ATP キット 製品一覧
  • 検出方法

    Colorimetric/Fluorometric
  • サンプルの種類

    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate
  • アッセイタイプ

    Quantitative
  • 検出感度

    < 1 µM
  • 全工程の試験時間

    1h 00m
  • 製品の概要

    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.

  • 特記事項

    Related assays

    Review the cell health assay guide to learn about kits to perform a cell viability assaycytotoxicity 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.

    How other researchers have used ATP Assay Kit ab83355

    This ATP assay kit has been used in publications in a variety of sample types, including:
    - Human: cell culture lysates1, primary monocyte cell culture lysates2, HCT116 cell culture supernatants3
    - Mouse: heart tissue4, liver5, C2C12 and L929 cell lysates6, primary thymocyte cell culture lysates7, cardiac tissue8
    - Rat: primary hippocampal neuron cell culture lysates9, liver tissue10, skeletal muscle11
    - Pig: kidney cell culture lysates12, heart tissue13 
    - C elegans tissue14
    - Chlamydomonas reinhardtii algae15

     

    References: 1 - Civallero M et al 2017, Na JY et al 2018, 2Gkirtzimanaki K et al 2018, 3Yang et al 2018; 4 - Singh SP et al 2018; 5 - Han SJ et al 2018; 6 - Alhindi Y et al 2019, Gregorczyk KP et al 2018; 7 - Simula L et al 2018; 8 - Litt MJ et al 2017, Samokhvalov V et al 2018; 9 - Zhao X et al 2018;  10 - Jing R et al 2018; 11 - Trinchese G et al 2018; 12 - Zou X et al 2018; 13 - Yuan F et al 2018; 14 - Pandey et al 2018; 15 - Ramanan R et al 2018

  • 試験プラットフォーム

    Microplate reader

製品の特性

画像

  • Ab83355 was used to determin ATP levels in rat pancreas islets as an ischemic marker to predict transplantation outcomes. We extracted ATP from fresh pancreas that have undergo different time of cold ischemia : 0, 2, 4, 6, 8 and 10h and 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.

  • Maiti AK et al (2018) used ATP assay kit ab83355 to measure mitochondrial ATP generation in an in vitro mouse intestinal model treated with cytokines in the presence and absence of VIP (vasoactive intestinal peptide). VIP was induced by C. rodentium infection and cytokines.

  • 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).

  • 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 10cells per well and incubated for two days under standard culture conditions. ATP levels are measured using ATP assay kit (ab83355).

  • Example of fluorometric ATP assay standard curve.
  • Maiti AK et al. (2015) used ATP assay kit ab113852 to measure mitochondrial ATP generation in murine distal colon after C. rodentium infection.

  • Example of colorimetric ATP assay standard curve.
  • 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).

プロトコール

参考文献

This product has been referenced in:

  • Alhindi Y  et al. Low Citrate Synthase Activity Is Associated with Glucose Intolerance and Lipotoxicity. J Nutr Metab 2019:8594825 (2019). Read more (PubMed: 30944739) »
  • Ahmed ME  et al. Synergy in Disruption of Mitochondrial Dynamics by Aß (1-42) and Glia Maturation Factor (GMF) in SH-SY5Y Cells Is Mediated Through Alterations in Fission and Fusion Proteins. Mol Neurobiol N/A:N/A (2019). Read more (PubMed: 30949973) »
See all 91 Publications for this product

レビューと Q&A

1-10 of 36 Abreviews or Q&A

Question
Answer

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

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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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Answer

This kit will detects total ATP.

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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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Answer

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

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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

Read More
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

投稿 Aug 07 2018

Testing ab83355 kit in cell culture media

Good Excellent 5/5 (Ease of Use)
Abreviews
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

投稿 Jul 31 2018

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.

Abcam user community

Verified customer

投稿 Mar 27 2018

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.

Mrs. Fotini Mouth

Verified customer

投稿 Nov 23 2017

1-10 of 36 Abreviews or Q&A

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