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) 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.

  • 特記事項

    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.

     

  • アプリケーション
    適用あり: Functional Studiesmore details

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab83355 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Functional Studies Use at an assay dependent dilution.

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  • 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.
  • 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:
  • Kanuri BN  et al. Altered glucose and lipid homeostasis in liver and adipose tissue pre-dispose inducible NOS knockout mice to insulin resistance. Sci Rep 7:41009 (2017). Functional Studies ; Mouse . Read more (PubMed: 28106120) »
  • Pushpakumar S  et al. Toll-like Receptor 4 Deficiency Reduces Oxidative Stress and Macrophage Mediated Inflammation in Hypertensive Kidney. Sci Rep 7:6349 (2017). Read more (PubMed: 28743964) »

See all 31 Publications for this product

レビューと Q&A

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

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.
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Mrs. Fotini MOUTH

Verified customer

投稿 Nov 23 2017

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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Thank you for contacting us at Abcam Scientific Support.

I am happy to assist with your protocol inquiries.

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

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Since there are enzymes in the assay which might need metal ions for function, we do not recommend metal chelators like EDTA for blood collection. Heparin is a better choice.


Generally, fresh samples are best, but frozen samples can be used with this kit. We suggest snap-freezing cells or tissue, and deproteinizing lysates before freezing, for best results.

Using a 561 nm excitation rather than the recommended 535 nm would compromise the results. Typically, bandpass filters are +/- 10nm, so you're 561 nm excitation would have a flexibility of 551 - 571 nm. If you do not have the means of exciting at 535 n...

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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.

This kit will detects total ATP.

Yes, you will have to use a homogenizer to ensure proper tissue lysis in the assay buffer. Any type of homogenizer that you prefer can be used, and as shown on the datasheet, we recommend deproteinizing sample isolation preferably with PCA rather than ...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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