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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. |
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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. Invitrogen Molecular Probes tools provide:
| U2OS cells were transduced with Prem Autophagy Sensor LC3B-RFP (red) and CellLight Tubulin-GFP (green). |
Invitrogen Premo Autophagy Sensor Tandem LC3B can be used to discriminate between acidic and neutral LC3B-positive vesicles.
| Premo Autophagy Sensor Tandem RFP-GFP-LC3B can be used to discriminate between acidic and neutral LC3B-positive vesicles. |
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. Invitrogen Molecular Probes tools enable:
| Imaging motility of p62-positive vesicles along microtubules using Premo Autophagy Sensor GFP-p62 Kit. |
Target | LC3B associates with the phagophore (nascent autophagosomal membrane) early in autophagy | ||
Readout | Localization of LC3B by fluorescence imaging | ||
Fluorescent label | GFP | RFP | Requires secondary detection |
Standard filter set | FITC | TRITC | NA |
Ex/Em (nm) | 485/520 | 555/584 | NA |
Signal-to-noise ratio | NA | ||
Photostability | NA | ||
Bibliography | Citations | Citations | |
Multiplexing | Yes | Yes | |
Live cells | Yes | No | |
Fixed cells | No | Yes | |
Fixable | Yes | Yes | |
Format | 1 kit | 1 kit | 1 kit |
Cat. No. | P36235 | P36236 | L10382 |
Target | LC3B associates with the phagophore (nascent autophagosomal membrane) early in autophagy and pH decreases after fusion with the lysosome | |
Readout | Tandem sensor to monitor LC3B progression from autophagosome to autolysosome | |
Fluorescent label | GFP | RFP |
Standard filter set | FITC | TRITC |
Ex/Em (nm) | 485/520 | 555/584 |
Signal-to-noise ratio | ||
Photostability | ||
Bibliography | Citations | |
Multiplexing | Yes | |
Live cells | Yes | |
Fixed cells | No | |
Fixable | No | |
Format | 1 kit | |
Cat. No. | P36239 |
Target | p62 binds to ubiquitin cargo and LC3 or its homologs | |
Readout | Localization of p62 by fluorescence imaging | |
Fluorescent label | GFP | RFP |
Standard filter set | FITC | TRITC |
Ex/Em (nm) | 485/520 | 555/584 |
Signal-to-noise ratio | ||
Photostability | ||
Bibliography | Citations | |
Multiplexing | Yes | |
Live cells | Yes | |
Fixed cells | No | |
Fixable | Yes | |
Format | 1 kit | 1 kit |
Cat. No. | P36240 | P36241 |
Target | LC3B associates with the phagophore (nascent autophagosomal membrane) early in autophagy | ||
Readout | Localization of LC3B by fluorescence imaging | ||
Fluorescent label | GFP | RFP | Requires secondary detection |
Standard filter set | FITC | TRITC | NA |
Ex/Em (nm) | 485/520 | 555/584 | NA |
Signal-to-noise ratio | NA | ||
Photostability | NA | ||
Bibliography | Citations | Citations | |
Multiplexing | Yes | Yes | |
Live cells | Yes | No | |
Fixed cells | No | Yes | |
Fixable | Yes | Yes | |
Format | 1 kit | 1 kit | 1 kit |
Cat. No. | P36235 | P36236 | L10382 |
Target | LC3B associates with the phagophore (nascent autophagosomal membrane) early in autophagy and pH decreases after fusion with the lysosome | |
Readout | Tandem sensor to monitor LC3B progression from autophagosome to autolysosome | |
Fluorescent label | GFP | RFP |
Standard filter set | FITC | TRITC |
Ex/Em (nm) | 485/520 | 555/584 |
Signal-to-noise ratio | ||
Photostability | ||
Bibliography | Citations | |
Multiplexing | Yes | |
Live cells | Yes | |
Fixed cells | No | |
Fixable | No | |
Format | 1 kit | |
Cat. No. | P36239 |
Target | p62 binds to ubiquitin cargo and LC3 or its homologs | |
Readout | Localization of p62 by fluorescence imaging | |
Fluorescent label | GFP | RFP |
Standard filter set | FITC | TRITC |
Ex/Em (nm) | 485/520 | 555/584 |
Signal-to-noise ratio | ||
Photostability | ||
Bibliography | Citations | |
Multiplexing | Yes | |
Live cells | Yes | |
Fixed cells | No | |
Fixable | Yes | |
Format | 1 kit | 1 kit |
Cat. No. | P36240 | P36241 |