Autophagy Assay试剂盒

产品详情

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.