High-Throughput Techniques for Autophagy Research

High-Throughput Techniques for Autophagy Research
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Aberrant autophagy is implicated in many important disease states such as cancer, neurodegeneration, and immunological diseases, and therefore small molecules that can remedy these deficiencies hold great therapeutic promise. The ability to study autophagosome formation and other aspects of autophagy, and to observe the effects of small molecules and gene manipulation on them in high throughput using physiologically relevant cell models, is of great importance, not only in understanding autophagy but also in producing effective treatments.

Autophagy describes the segregation and delivery of cytoplasmic cargo, including proteins and organelles, for degradation by hydrolytic enzymes through the lysosomal machinery. It is critical to the healthy functioning of cells and the failure of autophagy is a major reason for the accumulation of cell damage and aging.

Select Products for High-Throughput Techniques

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Locating and Tracking LC3B in Fixed Cells

The LC3 Antibody Kit for Autophagy includes a rabbit polyclonal antibody against LC3B that has been validated for use in fluorescence microscopy and high-content imaging and analysis.

  • Quantitative image analysis
  • Validated in high content screening applications
  • Autophagy disrupter included


See selection guide for Tracking LC3B for HCS

  Autophagy detected in HeLa cells using anti-LC3B
Autophagy detected in HeLa cells using anti-LC3B and Alexa Fluor® 488 goat anti–rabbit IgG.

Quantitative Assay for LC3B Accumulation

Measure the accumulation of autophagosome-associated LC3B-II in cell lysates using Tb-based TR-FRET

  • High-throughput analysis
  • LanthaScreen® format


See selection guide for Quantifying LC3B for HTS

  Quantifying autophagy with the Premo™ Autophagy TR-FRET LC3B Immunoassay
Quantifying autophagy with the Premo™ Autophagy TR-FRET LC3B Immunoassay.


Selection Guides for Products for High-Throughput Techniques

 
Target LC3B associates with the phagophore early in autophagy
Readout Localization of LC3B by fluorescence imaging
Fluorescent label Requires secondary detection
Bibliography Citations
Multiplexing Yes
Live cells No
Fixed cells Yes
Fixable Yes
Format 1 kit
Cat. No. L10382
 
Target protein LC3B associates with the phagophore early in autophagy
Readout Accumulation of LC3B-II measured in cell lysates using Tb-based TR-FRET
Fluorescent label GFP and Tb-labeled LC3B-II detection antibody
Standard filter set LanthaScreen®-specific instrument settings
Bibliography Citations
Multiplexing No
Live cells No
Fixed cells No
Fixable No
Format 1 kit (333 tests in 96-well format)
Cat. No. A14071

Resources

Molecular Probes® Handbook
BioProbes® Journal Article
Technical Resources
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