Name: ATP Assay Kit (Colorimetric/Fluorometric)
SKU: ab83355
Rating: 4 (5 reviews)

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Key features and details

Assay type: Quantitative
Detection method: Colorimetric/Fluorometric
Platform: Microplate reader
Assay time: 1 hr
Sample type: Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine (UTI)
Sensitivity: 1 µM

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

Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Urine (UTI)
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 ATP assay method. This kit can detect as low as 1 μM of ATP in various samples

How the assay works
This ATP assay kit uses the standard established assay protocol for fluorometric and colorimetric ATP determination kits: - glycerol is phosphorylated by glycerol kinase in an ATP dependent reaction to produce glycerol 3-phosphate - glycerol 3-phosphate is then acted on by glycerol phosphate oxidase to produce glycerone phosphate and hydrogen peroxide - hydrogen peroxide is then processed by a peroxidase causing a reaction with a probe to generate red color (λmax = 570 nm) and fluorescence (Ex/Em = 535/587 nm).

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

Related ATP assay products
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.

How other researchers are using 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}

Other notes
This product is manufactured by BioVision, an Abcam company and was previously called K354 ATP Colorimetric/Fluorometric Assay Kit. K354-100 is the same size as the 100 test size of ab83355.
特記事項

The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the SDS download section.
試験プラットフォーム

Microplate reader

保存方法

Store at -20°C. Please refer to protocols.

内容	100 tests	2000 tests
Assay Buffer XXIII	1 x 25ml	20 x 25ml
ATP Standard II	1 vial	20 vials
Converter Enzyme I	1 vial	20 vials
Developer IV	1 vial	20 vials
OxiRed Probe	1 x 200µl	20 x 200µl

研究分野
別名
- Adenosine 5' triphosphate

Corresponding kit
- Deproteinizing Sample Preparation Kit - TCA (ab204708)

ATP levels in Pancreatic IsletImage courtesy of Mrs. Fotini Mouth

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.
ATP assay used to study mitochondrial disfunction after C. rodentium infection in miceImage courtesy of Maiti A K et al. PLoS One. 2018; 13(9): e0204567. doi: 10.1371/journal.pone.0204567

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.
ATP assay performed with ab83355Sanokawa-Akakura R et al., PLoS One 9(9), fig4b. doi: 10.1371/journal.pone.0108537 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

The chart shows a comparison of ATP levels of HepG2 treated with 0, 10 and 100 µM for 48 hours, DR^{H2O2 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 ab83355Image from Shi Z et al., PLoS One 9(8), Fig 2 doi: 10.1371/journal.pone.0105675. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

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 10⁵cells 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

Example of fluorometric ATP assay standard curve.
ATP levels measured in mouse colon tissueImage courtesy of Maiti A K et al. Sci Rep. 2015; 5: 1543. doi: 10.1038/srep15434. Reproduced under the Creative Commons License http://creativecommons.org/licenses/by/4.0/

Maiti AK et al. (2015) used ATP assay kit ab113852 to measure mitochondrial ATP generation in murine distal colon after C. rodentium infection.
ATP assay performed with ab83355

Example of colorimetric ATP assay standard curve.
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).

Protocol Booklet

Click here to view the general protocols

SDS download

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ab83355 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab83355 は 435 報の論文で使用されています。

Peng ZM et al. PFKP deubiquitination and stabilization by USP5 activate aerobic glycolysis to promote triple-negative breast cancer progression. Breast Cancer Res 26:10 (2024). PubMed: 38217030
Mei Z et al. NAT10 mediated ac4C acetylation driven m6A modification via involvement of YTHDC1-LDHA/PFKM regulates glycolysis and promotes osteosarcoma. Cell Commun Signal 22:51 (2024). PubMed: 38233839
Mai N et al. Bioenergetic and excitotoxic determinants of cofilactin rod formation. J Neurochem 168:899-909 (2024). PubMed: 38299375
Bamgbose G & Tulin A PARP-1 is a transcriptional rheostat of metabolic and bivalent genes during development. Life Sci Alliance 7:N/A (2024). PubMed: 38012002
Gartz M et al. ACTA1 H40Y mutant iPSC-derived skeletal myocytes display mitochondrial defects in an in vitro model of nemaline myopathy. Exp Cell Res 424:113507 (2023). PubMed: 36796746

View all Publications for this product

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1-5 of 5 Abreviews

We tested the kit ab83355 to quantify the total ATP in 80 hours post fertilization zebrafish larvae. ATP was quantified by fluorometric method (Ex/Em = 535/587 nm) using a TECAN Infinite 200 PRO.
Ten l0 hours post fertilization (hpf) larvae were prepared following the protocol given by Abcam. Two technical replicates were run in parallel with a background control (mix without ATP converter).

According to our results, the background noise generate by glycerol phosphate does not allow the quantification of ATP in the larvae. Additionally, the kit seems to be not sensitive enough to detected ATP in 80 hpf zebrafish larvae. In fact, all measured values are below the last point of the calibration curve.

We tried different experimental set ups:
Fig. 1: Deproteinization using PCA (45 minutes) as described by the Abcam protocol. Each bar represents a biological replicate. 50 μl of sample was tested.
Fig. 2: Different deproteinization times (5 and 1 minute) and no deproteinization step. 25 μl of sample was tested.
Fig.3: Different dilutions (10 larvae in 100 μl buffer and 50 μl buffer) and clean-up without using acids (chloroform clean-up). 40 and 30 μl of sample were tested, respectively. Experiments were performed in duplicates.

Despite we tried different dilutions, deproteinization and clean-up steps, the general low concentration of ATP and the strong background noise does not allow the quantification of ATP in the larvae. Consequently, we do not recommend to use this kit for zebrafish larvae.
However, we would happy to try a more sensitive methods such as the Luminescent ATP Detection Assay Kit (ab113849).