Cell Meter™ Autophagy Assay Kit *Blue Fluorescence*
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Alternative formats
Cell Meter™ Autophagy Assay Kit *Green Fluorescence* |
Related products
Overview | ![]() ![]() |
Autophagy is an evolutionarily conserved degradation process that targets long-lived proteins, organelles, and other cytoplasmic components for degradation via the lysosomal pathway. The autophagy pathway is complementary to the action of the ubiquitin-proteasome pathway which typically degrades short-lived proteins. Activation of the autophagy pathway is required for multiple cellular roles, including survival during starvation, the clearance of intracellular components, development, and immunity. In the absence of stress, autophagy serves a house-keeping function, removing damaged organelles and cellular components preventing cytotoxic effects. Decreases and defects in autophagy have been implicated in multiple diseases, for example Huntingtons, Alzheimers, and Parkinsons. In terms of cancer development, autophagy seems to play multiple roles. Decreased or absent expression of certain autophagy proteins, such as Beclin-1 and Bif-1, increases tumor susceptibility in mice while the overexpression of these proteins can repress cancer cell growth. However, autophagy is critical for the survival of cancer cells within the nutrient poor and hypoxic core of solid tumors. Cell Meter™ Autophagy Kit employs Autophagy Blue™ as a specific autophagosome marker to analyze the activity of autophagy. The assay is optimized for direct detection of autophagy in both detached and attached cells. The kit provides all the essential components for the assay protocol. Cell Meter™ Autophagy Kit is suitable for fluorescence microscope, fluorescence microplate reader.
Platform
Fluorescence microscope
Excitation | DAPI channel |
Emission | DAPI channel |
Recommended plate | Black wall/clear bottom |
Fluorescence microplate reader
Excitation | 330 nm |
Emission | 520 nm |
Cutoff | 475 nm |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | Bottom read mode |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells with your test compounds at the density of 1 - 2 × 104 cells/well
- Add Autophagy Blue™ working solution
- Incubate at 37°C for 15 - 60 minutes
- Wash the cells with Wash Buffer
- Monitor the fluorescence increase at Ex/Em= 330/520 nm (Cutoff = 475 nm), fluorescence microscope with DAPI filter set
Important notes
Thaw all the components at room temperature before starting the experiment.
PREPARATION OF WORKING SOLUTION
Add 20 μL of 500X Autophagy Blue™ (Component A) into 10 mL of Stain Buffer (Component B) and mix well to make Autophagy Blue™ working solution. Protect from light. Note: 20 μL of 500X Autophagy Blue™ (Component A) is enough for one 96-well plate.
For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html
SAMPLE EXPERIMENTAL PROTOCOL
- Culture cells to a density optimal for autophagy induction according to your specific induction protocol (about 1 - 2 x 104 cells/well/96-well plate). At the same time, culture a non-induced negative control cell population at the same density as the induced population for every labeling condition.
- Remove medium.
- Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of Autophagy Blue™ working solution into each well.
- Incubate the cells in a 37°C, 5% CO2 incubator for 15 to 60 minutes. Note: The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment.
- Wash the cells with Wash Buffer (Component C) for 3 - 4 times, then add 100 µL Wash Buffer (Component C) to each well. Note: It is recommended to increase either the labeling concentration or the incubation time to allow the dye to accumulate if the cells do not appear to be sufficiently stained.
- Monitor the fluorescence intensity with a fluorescence microplate reader at Ex/Em = 330/520 nm (Cutoff = 475 nm), a fluorescence microscope with DAPI filter set.
Images

Figure 1. Autophagy Blue™ labeled vesicles are induced by starvation in HeLa cells using Cell Meter™ Autophagy Assay Kit. HeLa cells were incubated in a regular DMEM medium as control (A) or in a serum-depleted medium as autophagy treatment (B) for 16 hours. Both control cells and starved cells were incubated with Autophagy Blue™ working solution for 30 minutes in a 37 °C, 5% CO2 incubator, and then washed four times with wash buffer. Cells were imaged immediately under a fluorescence microscope with a DAPI channel. Autophagy is indicated by bright blue dot staining of autophagic vacuoles (B).
Citations
View all 13 citations: Citation Explorer
Mitochondrial-Targeted Antioxidant MitoQ-Mediated Autophagy: A Novel Strategy for Precise Radiation Protection
Authors: Bao, Xingting and Liu, Xiongxiong and Wu, Qingfeng and Ye, Fei and Shi, Zheng and Xu, Dan and Zhang, Jinhua and Dou, Zhihui and Huang, Guomin and Zhang, Hong and others,
Journal: Antioxidants (2023): 453
Authors: Bao, Xingting and Liu, Xiongxiong and Wu, Qingfeng and Ye, Fei and Shi, Zheng and Xu, Dan and Zhang, Jinhua and Dou, Zhihui and Huang, Guomin and Zhang, Hong and others,
Journal: Antioxidants (2023): 453
Cell cycle arrest is an important mechanism of action of compound Kushen injection in the prevention of colorectal cancer
Authors: Sun, Jie and Li, Mei and Lin, Tingru and Wang, Di and Chen, Jingyi and Zhang, Yu and Mu, Qing and Su, Huiting and Wu, Na and Liu, Aiyu and others,
Journal: Scientific reports (2022): 1--16
Authors: Sun, Jie and Li, Mei and Lin, Tingru and Wang, Di and Chen, Jingyi and Zhang, Yu and Mu, Qing and Su, Huiting and Wu, Na and Liu, Aiyu and others,
Journal: Scientific reports (2022): 1--16
GDF-15 Deficiency Reduces Autophagic Activity in Human Macrophages In Vitro and Decreases p62-Accumulation in Atherosclerotic Lesions in Mice
Authors: Heduschke, Aline and Ackermann, Kathrin and Wilhelm, Beate and Mey, Lilli and Bonaterra, Gabriel Alejandro and Kinscherf, Ralf and Schwarz, Anja
Journal: Cells (2021): 2346
Authors: Heduschke, Aline and Ackermann, Kathrin and Wilhelm, Beate and Mey, Lilli and Bonaterra, Gabriel Alejandro and Kinscherf, Ralf and Schwarz, Anja
Journal: Cells (2021): 2346
Effects of miR-34b/miR-892a upregulation and inhibition of ABCB1/ABCB4 on melatonin-induced apoptosis in VCR-resistant oral cancer cells
Authors: Hsieh, Ming-Ju and Lin, Chiao-Wen and Su, Shih-Chi and Reiter, Russel J and Chen, Andy Wei-Ge and Chen, Mu-Kuan and Yang, Shun-Fa
Journal: Molecular Therapy-Nucleic Acids (2020): 877--889
Authors: Hsieh, Ming-Ju and Lin, Chiao-Wen and Su, Shih-Chi and Reiter, Russel J and Chen, Andy Wei-Ge and Chen, Mu-Kuan and Yang, Shun-Fa
Journal: Molecular Therapy-Nucleic Acids (2020): 877--889
Decreased expression of LRA4, a key gene involved in rhamnose metabolism, caused up-regulated expression of the genes in this pathway and autophagy in Pichia pastoris
Authors: Jiao, Jian and Wang, Shuai and Tian, Hui and Xu, Xinxin and Zhang, Yuhong and Liu, Bo and Zhang, Wei
Journal: AMB Express (2020): 1--9
Authors: Jiao, Jian and Wang, Shuai and Tian, Hui and Xu, Xinxin and Zhang, Yuhong and Liu, Bo and Zhang, Wei
Journal: AMB Express (2020): 1--9
The inhibitory effect of TU-100 on hepatic stellate cell activation in the tumor microenvironment
Authors: Wada, Yuma and Tokuda, Kazunori and Morine, Yuji and Okikawa, Shohei and Yamashita, Shoko and Ikemoto, Tetsuya and Imura, Satoru and Saito, Yu and Yamada, Shinichiro and Shimada, Mitsuo
Journal: Oncotarget (2020): 4593
Authors: Wada, Yuma and Tokuda, Kazunori and Morine, Yuji and Okikawa, Shohei and Yamashita, Shoko and Ikemoto, Tetsuya and Imura, Satoru and Saito, Yu and Yamada, Shinichiro and Shimada, Mitsuo
Journal: Oncotarget (2020): 4593
Sonodynamic therapy inhibits palmitate-induced beta cell dysfunction via PINK1/Parkin-dependent mitophagy
Authors: Guo, Tian and Liu, Tianyang and Sun, Yun and Liu, Xianna and Xiong, Rongguo and Li, He and Li, Zhitao and Zhang, Zhiguo and Tian, Zhen and Tian, Ye
Journal: Cell death & disease (2019): 457
Authors: Guo, Tian and Liu, Tianyang and Sun, Yun and Liu, Xianna and Xiong, Rongguo and Li, He and Li, Zhitao and Zhang, Zhiguo and Tian, Zhen and Tian, Ye
Journal: Cell death & disease (2019): 457
Cell Cycle Arrest and Autophagy Induced by Compound Kushen Injection in sw620 and sw480 Colorectal Cancer Cells with p53 Mutation
Authors: Sun, Jie and Wang, Di and Zhang, Yu and Mu, Qing and Li, Mei and Hou, Xiaoli and Chen, Jingyi and Yu, Xin and Wang, Shaojie and Yu, Weidong
Journal: (2019)
Authors: Sun, Jie and Wang, Di and Zhang, Yu and Mu, Qing and Li, Mei and Hou, Xiaoli and Chen, Jingyi and Yu, Xin and Wang, Shaojie and Yu, Weidong
Journal: (2019)
Blue light-emitting diodes induce autophagy in colon cancer cells by Opsin 3
Authors: Yoshimoto, Toshiaki and Morine, Yuji and Takasu, Chie and Feng, Rui and Ikemoto, Tetsuya and Yoshikawa, Kozo and Iwahashi, Syuichi and Saito, Yu and Kashihara, Hideya and Akutagawa, Masatake and others,
Journal: Annals of gastroenterological surgery (2018): 154--161
Authors: Yoshimoto, Toshiaki and Morine, Yuji and Takasu, Chie and Feng, Rui and Ikemoto, Tetsuya and Yoshikawa, Kozo and Iwahashi, Syuichi and Saito, Yu and Kashihara, Hideya and Akutagawa, Masatake and others,
Journal: Annals of gastroenterological surgery (2018): 154--161
Methods for Measuring Autophagy Levels in Disease
Authors: Phadwal, Kanchan and Kurian, Dominic
Journal: (2017): 195--211
Authors: Phadwal, Kanchan and Kurian, Dominic
Journal: (2017): 195--211
References
View all 28 references: Citation Explorer
beta-Elemene induces apoptosis as well as protective autophagy in human non-small-cell lung cancer A549 cells
Authors: Liu J, Hu XJ, Jin B, Qu XJ, Hou KZ, Liu YP.
Journal: J Pharm Pharmacol (2012): 146
Authors: Liu J, Hu XJ, Jin B, Qu XJ, Hou KZ, Liu YP.
Journal: J Pharm Pharmacol (2012): 146
Tgf-beta1 induces autophagy and promotes apoptosis in renal tubular epithelial cells
Authors: Xu Y, Yang S, Huang J, Ruan S, Zheng Z, Lin J.
Journal: Int J Mol Med. (2012)
Authors: Xu Y, Yang S, Huang J, Ruan S, Zheng Z, Lin J.
Journal: Int J Mol Med. (2012)
High-Throughput Screening for AntiInfluenza A Virus Drugs and Study of the Mechanism of Procyanidin on Influenza A VirusInduced Autophagy
Authors: Dai J, Wang G, Li W, Zhang L, Yang J, Zhao X, Chen X, Xu Y, Li K.
Journal: J Biomol Screen. (2012)
Authors: Dai J, Wang G, Li W, Zhang L, Yang J, Zhao X, Chen X, Xu Y, Li K.
Journal: J Biomol Screen. (2012)
An autophagy inhibitor enhances the inhibition of cell proliferation
Authors: Yao F, Wang G, Wei W, Tu Y, Tong H, Sun S.
Journal: Mol Med Report (2012): 84
Authors: Yao F, Wang G, Wei W, Tu Y, Tong H, Sun S.
Journal: Mol Med Report (2012): 84
Inhibition of induced autophagy increases apoptosis of Nara-H cells
Authors: Nakamura O, Hitora T, Akisue T, Kawamoto T, Yamagami Y, Yamamoto T.
Journal: Int J Oncol (2011): 1545
Authors: Nakamura O, Hitora T, Akisue T, Kawamoto T, Yamagami Y, Yamamoto T.
Journal: Int J Oncol (2011): 1545
A new fluorescence-based assay for autophagy
Authors: Proikas-Cezanne T, Codogno P.
Journal: Chem Biol (2011): 940
Authors: Proikas-Cezanne T, Codogno P.
Journal: Chem Biol (2011): 940
Reactive oxygen species contribute to oridonin-induced apoptosis and autophagy in human cervical carcinoma HeLa cells
Authors: Zhang YH, Wu YL, Tashiro S, Onodera S, Ikejima T.
Journal: Acta Pharmacol Sin (2011): 1266
Authors: Zhang YH, Wu YL, Tashiro S, Onodera S, Ikejima T.
Journal: Acta Pharmacol Sin (2011): 1266
Salvianolic acid B inhibits autophagy and protects starving cardiac myocytes
Authors: Han X, Liu JX, Li XZ.
Journal: Acta Pharmacol Sin (2011): 38
Authors: Han X, Liu JX, Li XZ.
Journal: Acta Pharmacol Sin (2011): 38
Quantification of nonselective bulk autophagy in S. cerevisiae using Pgk1-GFP
Authors: Welter E, Thumm M, Krick R.
Journal: Autophagy (2010): 794
Authors: Welter E, Thumm M, Krick R.
Journal: Autophagy (2010): 794
Concomitant inhibition of AKT and autophagy is required for efficient cisplatin-induced apoptosis of metastatic skin carcinoma
Authors: Claerhout S, Verschooten L, Van Kelst S, De Vos R, Proby C, Agostinis P, Garmyn M.
Journal: Int J Cancer (2010): 2790
Authors: Claerhout S, Verschooten L, Van Kelst S, De Vos R, Proby C, Agostinis P, Garmyn M.
Journal: Int J Cancer (2010): 2790
Application notes
FAQ
Can I use any of your Cell Meter™ Autophagy Assay Kits with a flow cytometer?
How do cells protect themselves against cellular stress?
How does the autophagy marker in your Cell Meter™ Autophagy Assay Kit (Cat#23000) work?
What are the most common types of cell death?
Are Cell Navigator® Cell Plasma Membrane Staining Kits suitable for cell culture medium samples?
How do cells protect themselves against cellular stress?
How does the autophagy marker in your Cell Meter™ Autophagy Assay Kit (Cat#23000) work?
What are the most common types of cell death?
Are Cell Navigator® Cell Plasma Membrane Staining Kits suitable for cell culture medium samples?