LC3B & p62
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To enable autophagy studies, we offer specific tools to report LC3B and p62 localization. Labeling of p62 serves as a useful marker for the induction of autophagy, clearance of protein aggregates, and the inhibition of autophagy. Labeling of LC3B serve to track the binding of p62 and subsequent recruitment of autophagosomes. Premo Autophagy Sensor Tandem LC3B can be used to discriminate between acidic and neutral LC3B-positive vesicles providing a more in-depth investigation of autophagy in live cells, over time.

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 LC3B and p62 autophagy research products

Tracking Autophagy With LC3B

LC3B is recruited from the cytosol and associates with the phagophore early in autophagy. This localization serves as a general marker for autophagic membranes and for monitoring the process as it develops. Molecular Probes® tools provide:

  • Tracking and localization in live and fixed cells
  • pH monitoring in real time of vesicles containing LC3B
  • Quantitative assay for LC3B accumulation

See selection guide for Monitoring LC3B Recruitment

  Autophagy Research
U2OS cells were transduced with Premo™ Autophagy Sensor LC3B-RFP (red) and CellLight® Tubulin-GFP (green).

Monitoring Autophagosomal Maturation

Premo™ Autophagy Sensor Tandem LC3B can be used to discriminate between acidic and neutral LC3B-positive vesicles.

  • Monitors autophagy progression

 
See selection guide for Tracking LC3B Migration

  Autophagy Research
Premo™ Autophagy Sensor Tandem RFP-GFP-LC3B can be used to discriminate between acidic and neutral LC3B-positive vesicles.

Tracking Selective Autophagy With p62

p62 is a receptor for cargo destined to be degraded by autophagy, including ubiquitinated protein aggregates destined for clearance. The p62 protein is able to bind ubiquitin and also to LC3, thereby targeting the autophagosome and facilitating clearance of ubiquitinated proteins. Molecular Probes® tools enable:

  • Tracking p62 in live cells
  • Multiplexing with cellular markers


See selection guide for Targeting p62

  Autophagy Research
Imaging motility of p62-positive vesicles along microtubules using Premo™ Autophagy Sensor GFP-p62 Kit.


Selection Guides for LC3B and p62 Autophagy Research Products

 
TargetLC3B associates with the phagophore (nascent autophagosomal membrane) early in autophagy
ReadoutLocalization of LC3B by fluorescence imaging
Fluorescent labelGFPRFPRequires secondary detection
Standard filter setFITCTRITCNA
Ex/Em (nm)485/520 555/584 NA
Signal-to-noise ratioNA
PhotostabilityNA
BibliographyCitationsCitations
MultiplexingYesYes
Live cellsYesNo
Fixed cellsNoYes
FixableYesYes
Format1 kit1 kit1 kit
Cat. No.P36235P36236L10382
 
TargetLC3B associates with the phagophore (nascent autophagosomal membrane) early in autophagy and pH decreases after fusion with the lysosome
ReadoutTandem sensor to monitor LC3B progression from autophagosome to autolysosome
Fluorescent labelGFPRFP
Standard filter setFITCTRITC
Ex/Em (nm)485/520 555/584
Signal-to-noise ratio
Photostability
BibliographyCitations
MultiplexingYes
Live cellsYes
Fixed cellsNo
FixableNo
Format1 kit
Cat. No.P36239
 
Targetp62 binds to ubiquitin cargo and LC3 or its homologs
ReadoutLocalization of p62 by fluorescence imaging
Fluorescent labelGFPRFP
Standard filter setFITCTRITC
Ex/Em (nm)485/520 555/584
Signal-to-noise ratio
Photostability
BibliographyCitations
MultiplexingYes
Live cellsYes
Fixed cellsNo
FixableYes
Format1 kit1 kit
Cat. No.P36240P36241