Shop All Autophagy Detection Kits

Premo™ Autophagy Sensor LC3B-RFP (BacMam 2.0) (Invitrogen™)

Premo™ Autophagy Sensor combines the selectivity of a LC3B-red fluorescent protein (RFP) chimera with the transduction efficiency of BacMam technology, enabling unambiguous visualization of this protein. BacMam reagents (insect Baculovirus with a Mammalian promoter) are non-replicating in mammalian cells and thus safe to handle. They are also non-cytotoxic and ready-to-use. Unlike expression vectors, BacMam reagents enable titratable and reproducible expression and offer high co-transduction efficiency, therefore multiple BacMam reagents can readily be used in the same cell. Recent improvements made to the BacMam system enable efficient transduction in a wider variety of cells including neurons and neural stem cells (NSCs) with an easy, one-step protocol. Now, to visualize autophagy, simply add the BacMam LC3B-FP to the cells and incubate overnight for protein expression. Each Premo™ Autophagy Sensor Kit includes the BacMam LC3B-FP, a control BacMam LC3B (G120A)-RFP (mutation on the control BacMam LC3B prevents cleavage and subsequent lipidation during normal autophagy, thus protein localization should remain cytosolic and diffuse) and chloroquine diphosphate to artificially induce autophagosome accumulation.

Premo™ Autophagy Tandem Sensor RFP-GFP-LC3B Kit (Invitrogen™)

The Premo™ Autophagy Tandem Sensor RFP-GFP-LC3B Kit combines the ability to monitor the various stages of autophagy (through LC3B protein localization) with the high transduction efficiency and minimal toxicity of BacMam 2.0 expression technology. To perform image-based analysis for autophagy, simply add the BacMam 2.0 RFP-GFP-LC3B reagent to your mammalian cells, incubate overnight to ensure maximum protein expression, and then visualize using standard GFP (green fluorescent protein) and RFP (red fluorescent protein) settings.

This tandem RFP-GFP sensor capitalizes on the pH difference between the acidic autolysosome and the neutral autophagosome and the pH sensitivity differences exhibited by GFP (green fluorescent protein) and RFP (red fluorescent protein) to monitor progression from the autophagosome to autolysosome. The RFP and the GFP genes included in this chimera are TagRFP and Emerald GFP, respectively.

Each Premo™ Autophagy Tandem Sensor RFP-GFP-LC3B Kit includes BacMam 2.0 RFP-GFP-LC3B reagent and chloroquine diphosphate, which is used to inhibit autophagy.

Multiplex inspired: combining the Premo™ Autophagy Tandem Sensor with the far-red emitting LysoTracker® Deep Red (available separately) allows for a three-color analysis of the complete autophagic pathway dynamics
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

The LC3B protein plays a critical role in autophagy, and the localization of this protein to autophagosomes can be used as a general marker for formation of autophagosomes during stimulation of autophagy and/or the accumulation of autophagosomes during inhibition of autophagic flux.

Currently available non-tandem autophagy chimeras, such as Premo™ Autophagy LC3B-GFP (P36235) and LC3B-RFP (P36236), are useful for monitoring autophagosome formation, especially when combined with other chimeras or fluorescent reagents. However, the Premo™ Autophagy Tandem Sensor allows an enhanced dissection of the maturation of the autophagosome to the autolysosome. By combining an acid-sensitive GFP with an acid-insensitive RFP, the change from autophagosome (neutral pH) to autolysosome (with an acidic pH) can be visualized by imaging the specific loss of the GFP fluorescence, leaving only red fluorescence. In addition, combining the Premo™ Autophagy Tandem Sensor with the far-red emitting LysoTracker® Deep Red (L12492) allows for a three-color analysis of the autophagosomal/autolysosomal/lysosomal dynamics.

To ensure reliable, high levels of expression in a wide variety of mammalian cells, the Premo™ Autophagy Sensors utilize BacMam 2.0 expression 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 Tandem Sensor RFP-GFP-LC3B 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.

Premo™ Autophagy Sensor RFP-p62 Kit (Invitrogen™)

The Premo™ Autophagy Sensor RFP-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 RFP-p62 reagent to your mammalian cells, incubate overnight to ensure adequate protein expression, and then visualize using standard RFP (red fluorescent protein) settings. The RFP gene included in this chimera is mKate2. This monomeric fluorescent protein is very bright, has excellent pH resistance, and improved photostability.

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

This RFP-p62 chimera is recommended for use with green-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 RFP-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.

LC3B Antibody Kit for Autophagy (Invitrogen™)

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. Chloroquine diphosphate is also included - following treatment with this compound, normal autophagic flux is disrupted and autophagosomes accumulate as a result of the disruption in normal autophagic flux.

Premo™ Autophagy Sensor LC3B-GFP (BacMam 2.0) (Invitrogen™)

Premo™ Autophagy Sensor combines the selectivity of a LC3B-green fluorescent protein (GFP) chimera with the transduction efficiency of BacMam technology, enabling unambiguous visualization of this protein. BacMam reagents (insect Baculovirus with a Mammalian promoter) are non-replicating in mammalian cells and thus safe to handle. They are also non-cytotoxic and ready-to-use. Unlike expression vectors, BacMam reagents enable titratable and reproducible expression and offer high co-transduction efficiency, therefore multiple BacMam reagents can readily be used in the same cell. Recent improvements made to the BacMam system enable efficient transduction in a wider variety of cells including neurons and neural stem cells (NSCs) with an easy, one-step protocol. Now, to visualize autophagy, simply add the BacMam LC3B-GFP to the cells and incubate overnight for protein expression. Each Premo™ Autophagy Sensor Kit includes the BacMam LC3B-FP, a control BacMam LC3B (G120A)-FP (mutation on the control BacMam LC3B prevents cleavage and subsequent lipidation during normal autophagy, thus protein localization should remain cytosolic and diffuse) and chloroquine diphosphate to artificially induce autophagosome accumulation.

Premo™ Autophagy Sensor GFP-p62 Kit (Invitrogen™)

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.