AB139484

Autophagy Assay Kit

Name: Autophagy Assay Kit (ab139484) | Abcam
Brand: Abcam Limited
SKU: AB139484
Price: 103000 JPY
Availability: InStock
Rating: 4 (2 reviews)

(2 Reviews)

(106 Publications)

Autophagy Assay Kit ab139484 measures autophagic vacuoles and monitors autophagic flux in live cells using a dye that selectively labels autophagic vacuoles.

- Single 30-min staining incubation with dye
- Readout with FITC channels/ filters on flow cytometer, fluorescent microscope, or fluorometric plate reader

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Functional Studies - Autophagy Assay Kit (AB139484)

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Functional Studies - Autophagy Assay Kit (AB139484)

Fluorescent microscopy analysis showing nucleus (blue nuclear stain; DAPI filter) and autophagic vesicules (green, FITC filter) in control HepG2 cells or cells starved in serum free medium for 5 hours to induce the formation of autophagic vesicles. HepG2 cells were also treated with 0.1 mM chloroquine for 24 hours or starved and chloroquine treated for 5 hours (in earlier stages of autophagy, chloroquine induces autophagosome formation).

FuncS

Lab

Functional Studies - Autophagy Assay Kit (AB139484)

Fluorescent microscopy analysis showing nucleus (blue nuclear stain; DAPI filter) and autophagic vesicules (green, FITC filter) in control HepG2 cells or cells treated with 1 uM Rapamycin (ab120224) for 24 hours.

Flow Cytometry - Autophagy Assay Kit (AB139484)

Flow Cyt

Supplier Data

Flow Cytometry - Autophagy Assay Kit (AB139484)

Jurkat cells (acute T-Cell leukemia), uninduced or treated overnight with 0.5 µM Rapamycin (a typical autophagy inducer) were loaded with Green Detection Reagent, then washed and analyzed by flow cytometry. Results are presented as histogram overlay. Control cells (blue solid line) were stained as well but mostly display low fluorescence. In the samples treated with 500 nM Rapamycin for 18 hours (black solid line), Green dye signal increases about 2-fold, indicating that Rapamycin induced autophagy in Jurkat cells.

Key facts

検出方法

Fluorescent

サンプルタイプ

Suspension cells, Adherent cells

アッセイタイプ

Cell-based

アッセイ時間

30m

アッセイプラットフォーム

Microplate reader, Fluor. microscope, Flow cyt.

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製品の詳細

Autophagy Assay Kit ab139484 offers a rapid and quantitative approach to monitoring autophagy in live cells without the need for cell transfection.It can be analyzed using flow cytometry, fluorescent microscopy, or using a microplate reader. The dye has spectral characteristics similar to FITC, and can be read with FITC channels / filters.

How the assay works
The Green Detection Reagent dye used in the Autophagy assay kit protocol has been optimized by screening dyes for both minimal staining of lysosomes, and bright fluorescence upon incorporation into pre-autophagosomes, autophagosomes, and autolysosomes (autophagolysosomes).

Autophagy inducer rapamycin is included in the kit as a positive control. Please note that the optimal final concentration is cell-dependent and should be determined experimentally for each cell line being tested. The agent has been validated in HeLa, HepG2 and Jurkat cells.

Chloroquine is included in the kit to use as an inhibitor control. Chloroquine may be used in combination with rapamycin or starvation in monitoring autophagic flux. Depending on the applications and specific cell lines, a 10-120 μM final concentration is recommended in order to observe changes in autophagic flux.

Autophagy assay protocol summary
- remove growth medium from cells
- add staining mix and incubate for 30 min
- wash cells
- analyze with fluorescence microscopy, flow cytometry, or fluorescent microplate reader

The reagents provided in this kit are sufficient for 200 flow cytometry tests, 250 fluorescence microscopy test or 3 x 96-well microplate tests.

How other researchers are using ab139484
Autophagy Assay Kit has been used in a variety of sample types including:
- Human normal hepatocytes and Human HCC cell lines 1
- Retinal endothelial cells, isolated from nondiabetic human retina 2
- 3T3-L1-adipocytes 3
References: 1 - Rajan P et al. 2023; 2 - Kowluru R et al. 2023; 3 - Park J at al. 2024.

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製品内容

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出荷温度及び保存条件

出荷温度

Dry Ice

短期保存温度

-80°C

長期保存温度

-80°C

保管に関する情報

-80°C

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補足情報

This supplementary information is collated from multiple sources and compiled automatically.

Autophagy also known as "self-eating" is a cellular degradation and recycling process critical for maintaining cellular homeostasis. Mechanically autophagy involves the formation of double-membrane vesicles called autophagosomes which engulf damaged organelles and proteins. The autophagosomes then fuse with lysosomes leading to the degradation of the contents by lysosomal enzymes. Autophagy is expressed highly in cells under stress such as nutrient deprivation and is a conserved process across eukaryotic cells. Its machinery involves more than 30 autophagy-related genes (ATGs) but does not focus on a single mass or protein as it is a complex pathway.

Biological function summary

Autophagy protects cells by degrading and recycling components therefore preventing accumulation of damaged proteins and organelles. It forms part of the cellular defense mechanisms against stress and aging contributing to cellular longevity. In starvation conditions autophagy provides an internal source of nutrients helping cell survival. The process is part of a larger complex involving ATG proteins which drive the sequential steps of autophagosome formation. Monodansylcadaverine an autofluorescent compound often marks autophagic vacuoles in experimental settings providing a tool for autophagy detection and study.

Pathways

Autophagy is deeply integrated into cellular signaling networks. It plays a significant role in the mTOR (mechanistic target of rapamycin) signaling pathway which senses nutrient availability and regulates cell growth. Autophagy also intersects with the AMPK (AMP-activated protein kinase) pathway which responds to energy stress promoting catabolism when cellular ATP levels drop. These intersections with mTOR and AMPK pathways illustrate autophagy's essential role in balancing anabolic and catabolic processes and its regulatory association with proteins involved in cellular stress responses like p62/SQSTM1.

Autophagy is relevant to conditions like cancer and neurodegeneration. In cancer autophagy can have dual roles both supporting tumor cell survival under metabolic stress and limiting unregulated cell division. The Bcl-2 protein family which regulates apoptosis also modulates autophagy highlighting a complex interaction between cell death and survival. In neurodegenerative diseases such as Alzheimer's impaired autophagy leads to the accumulation of protein aggregates contributing to neuronal damage. Here proteins linked to autophagic dysfunction include beta-amyloid and tau both of which are involved in Alzheimer's disease pathology.

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製品プロトコール

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ターゲットの情報

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文献 (106)

Recent publications for all applications. Explore the full list and refine your search

Nutrients 17: PubMed40290068

2025

l-Carnitine and Acetyl-l-Carnitine Induce Metabolism Alteration and Mitophagy-Related Cell Death in Colorectal Cancer Cells.

Applications

Unspecified application

Species

Unspecified reactive species

Isabella Donisi,Anna Balestrieri,Vitale Del Vecchio,Giovanna Bifulco,Maria Luisa Balestrieri,Giuseppe Campanile,Nunzia D'Onofrio

Journal of cellular and molecular medicine 29:e70528 PubMed40205749

2025

Ambrisentan Retains Its Pro-Autophagic Activity on Human Pulmonary Artery Endothelial Cells Exposed to Hypoxia in an In Vitro Model Mimicking Diabetes.

Applications

Unspecified application

Species

Unspecified reactive species

Manuela Cabiati,Filippo Biondi,Sandra Ghelardoni,Valentina Casieri,Vincenzo Lionetti,Agnese Sgalippa,Silvia Del Ry,Rosalinda Madonna

Metabolites 15: PubMed40137158

2025

Modulation of Peripheral Mast Cell and Brain Microglia Axis via Kinase Inhibition.

Applications

Unspecified application

Species

Unspecified reactive species

Xiaoguang Liu,Michaeline Hebron,Kaluvu Balaraman,Louis Ballard,Kimberly Liu,Max Stevenson,Charbel Moussa

Nucleic acids research 53: PubMed40105243

2025

DNA damage response regulator ATR licenses PINK1-mediated mitophagy.

Applications

Unspecified application

Species

Unspecified reactive species

Christian Marx,Xiaobing Qing,Yamin Gong,Joanna Kirkpatrick,Kanstantsin Siniuk,Galina V Beznoussenko,Gururaj Rao Kidiyoor,Murat Kirtay,Katrin Buder,Philipp Koch,Martin Westermann,Christopher Bruhn,Eric J Brown,Xingzhi Xu,Marco Foiani,Zhao-Qi Wang

BioMed research international 2025:5556353 PubMed40041570

2025

Expression of UCOE and HSP27 Molecular Elements to Improve the Stable Protein Production on HEK293 Cells.

Applications

Unspecified application

Species

Unspecified reactive species

Concepción Sosa-García,Uriel Abdallah Sánchez-Pacheco,Carlos Alberto Tavira-Montalvan,Angélica Meneses-Acosta

Cellular & molecular immunology 22:260-281 PubMed39910335

2025

Cell-associated galectin 9 interacts with cytotoxic T cells confers resistance to tumor killing in nasopharyngeal carcinoma through autophagy activation.

Applications

Unspecified application

Species

Unspecified reactive species

Ngar-Woon Kam,Cho Yiu Lau,Jeffrey Yan Ho Lau,Xin Dai,Yusi Liang,Syrus Pak Hei Lai,Michael King Yung Chung,Valen Zhuoyou Yu,Wenting Qiu,Mengsu Yang,Corey Smith,Rajiv Khanna,Kwan Ming Ng,Wei Dai,Chi Ming Che,Victor Ho-Fun Lee,Dora L W Kwong

Molecular microbiology 123:1-15 PubMed39626091

2024

Uncovering the Significance of JNK/AKT Axis in the Autophagic Regulation of Leishmania major Infection.

Applications

Unspecified application

Species

Unspecified reactive species

Vrushali Guhe,Shailza Singh

Journal of medicinal chemistry 67:19475-19502 PubMed39496093

2024

Optimization of Antiproliferative Properties of Triimine Copper(II) Complexes.

Applications

Unspecified application

Species

Unspecified reactive species

Katarzyna Choroba,Bartosz Zowiślok,Sławomir Kula,Barbara Machura,Anna M Maroń,Karol Erfurt,Cristiana Marques,Sandra Cordeiro,Pedro V Baptista,Alexandra R Fernandes

Aging biology 2: PubMed39263528

2024

Ezetimibe Lowers Risk of Alzheimer's and Related Dementias over Sevenfold, Reducing Aggregation in Model Systems by Inhibiting 14-3-3G::Hexokinase Interaction.

Applications

Unspecified application

Species

Unspecified reactive species

Akshatha Ganne,Nirjal Mainali,Meenakshisundaram Balasubramaniam,Ramani Atluri,Sonu Pahal,Joseph Asante,Corey Nagel,Srikanth Vallurupalli,Robert J Shmookler Reis,Srinivas Ayyadevara

Dalton transactions (Cambridge, England : 2003) 53:17803-17818 PubMed39233530

2024

Effect of mono- and dinuclear thiosemicarbazone platinacycles in the proliferation of a colorectal carcinoma cell line.

Applications

Unspecified application

Species

Unspecified reactive species

Francisco Reigosa-Chamorro,Sandra Cordeiro,M Teresa Pereira,Beatriz Filipe,Pedro V Baptista,Alexandra R Fernandes,José M Vila

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Autophagy Assay Kit