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Premo™ Autophagy Sensor GFP-p62 Kit

Catalog number: P36240

Molecular Probes™  Related applications: Cell Viability, Proliferation & Function

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Description

The Premo™ Autophagy Sensor GFP-p62 Kit combines the ability to monitor the induction or inhibition of autophagy, through the localization of the autophagy receptor p62 (also known as SQSTM1), with the high transduction efficiency and minimal toxicity of BacMam 2.0 technology. To perform an image-based analysis for autophagy, simply add the BacMam 2.0 GFP-p62 reagent to your mammalian cells, incubate overnight to ensure adequate protein expression, and then visualize using standard GFP (green fluorescent protein) settings. The GFP gene included in this chimera is TagGFP2, which has been demonstrated to mature 1.6-times faster than TagGFP and has increased pH stability when compared to Emerald GFP.

Each Premo™ Autophagy Sensor GFP-p62 Kit includes BacMam 2.0 GFP-p62 reagent and chloroquine diphosphate, which is used to inhibit autophagy.

This GFP-p62 chimera is recommended for use with red-emitting fluorescent proteins or dyes.

Highly efficient delivery system: >90% transduction of a wide range of mammalian cell lines, including primary cells, stem cells, and neurons
Fast and convenient: simply add the Premo™ Autophagy reagent to your cells, incubate overnight, and image—or store frozen, assay-ready cells for later use
Robust: the Premo™ Autophagy reagents are non-replicating in mammalian cells, lack observable cytopathic effect, and are suitable for biosafety level (BSL) 1
Multiplex-enabled: additional BacMam or fluorescent reagents can be used in conjugation with the Premo™ Autophagy products to detect and monitor additional targets of interest

The p62 protein, also known as sequestosome (SQSTM1), is an ubitiquitin-binding protein that functions as a receptor for cargos destined to be degraded by the cellular autophagic machinery. When autophagy is induced the p62 protein localizes to the autophagosomes and is subsequently degraded. Conversely, with the inhibition of autophagy, the p62 protein accumulates in the autophagosome. Thus, the subcellular localization a p62-fluorescent protein chimera serves as a useful marker for the induction and inhibition of autophagy.

To ensure reliable, high levels of expression in a wide variety of mammalian cells, the Premo™ Autophagy Sensors utilize BacMam 2.0 technology. BacMam reagents (insect Baculovirus with a Mammalian promoter) do not replicate in mammalian cells, are non-cytoxic (biosafety level 1), and are ready to use. Unlike expression vectors, BacMam reagents enable titratable and reproducible expression and offer high co-transduction efficiency; therefore, multiple BacMam reagents can be used in the same cell.

Each Premo™ Autophagy Sensor GFP-p62 Kit includes a vial of chloroquine diphosphate. Chloroquine has been demonstrated to inhibit autophagy by elevating lysosomal pH and therefore inhibiting the fusion of autophagosomes with lysosomes and preventing the subsequent lysosomal protein degradation.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Form: Liquid
Color: Green
Format: Tube(s)
System: Premo™
Label or Dye: GFP (TagGFP)
Product Size: 1 kit
Emission Class: Visible
Compatible Cells: Mammalian Cells
Detection Method: Fluorescent
Excitation Class: Visible
For Use With (Equipment): Fluorescence Microscope, Fluorescent Imager
Excitation⁄Emission (nm): 483⁄506

Contents & storage

Store at 2 to 6°C

DO NOT FREEZE

Protect material from long-term exposure to light, but may be exposed to light for short periods of time.

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Manuals & protocols

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