Invitrogen™

Premo Autophagy Assays with Sensor LC3B, p62, or Tandem Sensor RFP-GFP-LC3B

Catalog number: P36235
Invitrogen™

Premo Autophagy Assays with Sensor LC3B, p62, or Tandem Sensor RFP-GFP-LC3B

Catalog number: P36235
Easily monitor various stages of cellular autophagy with Premo Autophagy assays and sensors, which are based on LC3B- or p62-linked fluorescent protein chimeras, or the RFP-GFP-LC3B tandem sensor. Premo Autophagy assay kits offer the selectivity of Premo Autophagy sensors, which are based on chimeras of autophagy-associated proteins LC3B or p62, and green (GFP) and/or red fluorescent protein (RFP), with the transduction efficiency of BacMam technology. Transduction or even co-transduction can be performed in a wide variety of cells, including neurons and neural stem cells (NSCs) using a simple protocol that involves adding the BacMam 2.0 chimeric reagent to cells, incubating overnight, and then visualizing transduction using standard GFP (green fluorescent protein) and/or RFP (red fluorescent protein) settings.
 
Catalog Number
P36235
Unit Size
1 kit
Color
Green
Label or Dye
LC3B-GFP
Description
Premo Autophagy Sensor LC3B-GFP (BacMam 2.0)
Price (USD)
Price: 832.00
Your Price:
Availability
-
Full specifications
Quantity
DescriptionPremo Autophagy Sensor LC3B-GFP (BacMam 2.0)
FormatSlide
Kit ContentsLC3B-FP, a control BacMam LC3B (G120A)-RFP, and chloroquine diphosphate
Product TypeAutophagy Sensor
Compatible CellsMammalian Cells
Excitation/Emission488/520
ColorGreen
Shipping ConditionWet Ice
Product LinePremo
Detection MethodFluorescent
For Use With (Equipment)Confocal Microscope
Label or DyeLC3B-GFP
Label TypeOther Label(s) or Dye(s)
Storage RequirementsStore at 2°C to 6°C, protected from light. Do Not Freeze.
Unit Size1 kit
Catalog Number
Unit Size
1 kit
Color
Red
Label or Dye
LC3B-RFP
Description
Premo Autophagy Sensor LC3B-RFP (BacMam 2.0)
Price (USD)
Price: 806.00
Your Price:
Availability
-
Full specifications
Quantity
DescriptionPremo Autophagy Sensor LC3B-RFP (BacMam 2.0)
FormatSlide
Kit ContentsBacMam LC3B-FP, a control BacMam LC3B (G120A)-RFP, and chloroquine diphosphate
Product TypeAutophagy Sensor
Compatible CellsMammalian Cells
Excitation/Emission555/584
ColorRed
Shipping ConditionWet Ice
Product LinePremo
Detection MethodFluorescent
For Use With (Equipment)Confocal Microscope
Label or DyeLC3B-RFP
Label TypeOther Label(s) or Dye(s)
Storage RequirementsStore at 2°C to 6°C, protected from light. Do Not Freeze.
Unit Size1 kit
Catalog Number
Unit Size
1 kit
Color
Red-Orange, Green
Label or Dye
RFP-GFP-LC3B
Description
Premo Autophagy Tandem Sensor RFP-GFP-LC3B Kit
Price (USD)
Price: 842.00
Your Price:
Availability
-
Full specifications
Quantity
DescriptionPremo Autophagy Tandem Sensor RFP-GFP-LC3B Kit
FormatTube
Kit ContentsAutophagy Sensor RFP-GFP-LC3B, and chloroquine diphosphate
Product TypeAutophagy Sensor
Compatible CellsMammalian Cells
Excitation/EmissionTagRFP: 555/584
EmGFP: 488/509
ColorRed-Orange, Green
Shipping ConditionWet Ice
Product LinePremo
Detection MethodFluorescent
For Use With (Equipment)Fluorescence Microscope
Label or DyeRFP-GFP-LC3B
Label TypeOther Label(s) or Dye(s)
Storage RequirementsStore at 2°C to 6°C, protected from light. Do Not Freeze.
Unit Size1 kit
Catalog Number
Unit Size
1 kit
Color
Red
Label or Dye
RFP-p62
Description
Premo Autophagy Sensor RFP-p62 Kit
Price (USD)
Price: 792.00
Your Price:
Availability
-
Full specifications
Quantity
DescriptionPremo Autophagy Sensor RFP-p62 Kit
FormatTube
Kit ContentsAutophagy Sensor p62-RFP, and chloroquine diphosphate
Product TypeAutophagy Sensor
Compatible CellsMammalian Cells
Excitation/Emission588/633
ColorRed
Shipping ConditionWet Ice
Product LinePremo
Detection MethodFluorescent
For Use With (Equipment)Fluorescence Microscope
Label or DyeRFP-p62
Label TypeOther Label(s) or Dye(s)
Storage RequirementsStore at 2°C to 6°C, protected from light. Do Not Freeze.
Unit Size1 kit
Catalog Number
Unit Size
1 kit
Color
Green
Label or Dye
GFP-p62
Description
Premo Autophagy Sensor GFP-p62 Kit
Price (USD)
Price: 788.00
Your Price:
Availability
-
Full specifications
Quantity
DescriptionPremo Autophagy Sensor GFP-p62 Kit
FormatTube
Kit ContentsAutophagy Sensor p62-GFP, and chloroquine diphosphate
Product TypeAutophagy Sensor
Compatible CellsMammalian Cells
Excitation/Emission488/506
ColorGreen
Shipping ConditionWet Ice
Product LinePremo
Detection MethodFluorescent
For Use With (Equipment)Fluorescence Microscope
Label or DyeGFP-p62
Label TypeOther Label(s) or Dye(s)
Storage RequirementsStore at 2°C to 6°C, protected from light. Do Not Freeze.
Unit Size1 kit
Showing 5 of 5
Catalog NumberSpecificationsUnit SizeColorLabel or DyeDescriptionPrice (USD)AvailabilityQuantity
P36235Full specifications
1 kitGreenLC3B-GFPPremo Autophagy Sensor LC3B-GFP (BacMam 2.0)
Price: 832.00
Your Price:
-
DescriptionPremo Autophagy Sensor LC3B-GFP (BacMam 2.0)
FormatSlide
Kit ContentsLC3B-FP, a control BacMam LC3B (G120A)-RFP, and chloroquine diphosphate
Product TypeAutophagy Sensor
Compatible CellsMammalian Cells
Excitation/Emission488/520
ColorGreen
Shipping ConditionWet Ice
Product LinePremo
Detection MethodFluorescent
For Use With (Equipment)Confocal Microscope
Label or DyeLC3B-GFP
Label TypeOther Label(s) or Dye(s)
Storage RequirementsStore at 2°C to 6°C, protected from light. Do Not Freeze.
Unit Size1 kit
P36236Full specifications
1 kitRedLC3B-RFPPremo Autophagy Sensor LC3B-RFP (BacMam 2.0)
Price: 806.00
Your Price:
-
DescriptionPremo Autophagy Sensor LC3B-RFP (BacMam 2.0)
FormatSlide
Kit ContentsBacMam LC3B-FP, a control BacMam LC3B (G120A)-RFP, and chloroquine diphosphate
Product TypeAutophagy Sensor
Compatible CellsMammalian Cells
Excitation/Emission555/584
ColorRed
Shipping ConditionWet Ice
Product LinePremo
Detection MethodFluorescent
For Use With (Equipment)Confocal Microscope
Label or DyeLC3B-RFP
Label TypeOther Label(s) or Dye(s)
Storage RequirementsStore at 2°C to 6°C, protected from light. Do Not Freeze.
Unit Size1 kit
P36239Full specifications
1 kitRed-Orange, GreenRFP-GFP-LC3BPremo Autophagy Tandem Sensor RFP-GFP-LC3B Kit
Price: 842.00
Your Price:
-
DescriptionPremo Autophagy Tandem Sensor RFP-GFP-LC3B Kit
FormatTube
Kit ContentsAutophagy Sensor RFP-GFP-LC3B, and chloroquine diphosphate
Product TypeAutophagy Sensor
Compatible CellsMammalian Cells
Excitation/EmissionTagRFP: 555/584
EmGFP: 488/509
ColorRed-Orange, Green
Shipping ConditionWet Ice
Product LinePremo
Detection MethodFluorescent
For Use With (Equipment)Fluorescence Microscope
Label or DyeRFP-GFP-LC3B
Label TypeOther Label(s) or Dye(s)
Storage RequirementsStore at 2°C to 6°C, protected from light. Do Not Freeze.
Unit Size1 kit
P36241Full specifications
1 kitRedRFP-p62Premo Autophagy Sensor RFP-p62 Kit
Price: 792.00
Your Price:
-
DescriptionPremo Autophagy Sensor RFP-p62 Kit
FormatTube
Kit ContentsAutophagy Sensor p62-RFP, and chloroquine diphosphate
Product TypeAutophagy Sensor
Compatible CellsMammalian Cells
Excitation/Emission588/633
ColorRed
Shipping ConditionWet Ice
Product LinePremo
Detection MethodFluorescent
For Use With (Equipment)Fluorescence Microscope
Label or DyeRFP-p62
Label TypeOther Label(s) or Dye(s)
Storage RequirementsStore at 2°C to 6°C, protected from light. Do Not Freeze.
Unit Size1 kit
P36240Full specifications
1 kitGreenGFP-p62Premo Autophagy Sensor GFP-p62 Kit
Price: 788.00
Your Price:
-
DescriptionPremo Autophagy Sensor GFP-p62 Kit
FormatTube
Kit ContentsAutophagy Sensor p62-GFP, and chloroquine diphosphate
Product TypeAutophagy Sensor
Compatible CellsMammalian Cells
Excitation/Emission488/506
ColorGreen
Shipping ConditionWet Ice
Product LinePremo
Detection MethodFluorescent
For Use With (Equipment)Fluorescence Microscope
Label or DyeGFP-p62
Label TypeOther Label(s) or Dye(s)
Storage RequirementsStore at 2°C to 6°C, protected from light. Do Not Freeze.
Unit Size1 kit
Showing 5 of 5
Premo Autophagy sensors take advantage of autophagy-associated proteins LC3B or p62, along with GFP or RFP, to detect either the induction or inhibition of autophagy in cells. The Premo Autophagy sensors are transduced using BacMam (insect Baculovirus with a Mammalian promoter) technology, enabling unambiguous visualization of the chimeric proteins in autophagosomes. BacMam reagents are non-replicating in mammalian cells and therefore safe to handle. They are also non-cytotoxic and ready-to-use. Unlike typical expression vectors, BacMam reagents enable titratable and reproducible expression, and offer high co-transduction efficiency; therefore, multiple BacMam reagents can be readily used on the same cells. 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. To visualize autophagy, simply add the BacMam Autophagy Sensor to cells and incubate overnight for protein expression, then detect signal using standard GFP and/or RFP fluorescence settings on your instrument.

The LC3B protein plays a critical role in autophagy. Normally, this protein resides in the cytosol, but following cleavage and lipidation with phosphatidylethanolamine, LC3B associates with the phagophore. This localization can be used as a general marker for autophagic membranes. 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 of a p62-fluorescent protein chimera serves as a useful marker for the induction and inhibition of autophagy.

Additionally, by combining the acid-sensitive GFP (i.e., Emerald GFP) with an acid-insensitive RFP (i.e., TagRFP) in the tandem sensor RFP-GFP-LC3B, the change from an autophagosome (neutral pH) to the autolysosome (with an acidic pH) can be visualized by imaging the specific loss of the GFP fluorescence upon acidification of the autophagosome following lysosomal fusion. Upon induction of autophagy, the Premo Tandem Autophagy Sensor labels the punctate autophagosomes; these structures are positive for both GFP and RFP. Once the lysosome has fused, the pH drops, which quenches the GFP, making autolysosomes appear red. Additionally, combining the Premo Autophagy Tandem Sensor with the far-red emitting LysoTracker Deep Red (available separately) allows for a three-color analysis of autophagosomal/autolysomal/lysosomal dynamics.

Each Premo Autophagy assay kit includes chloroquine diphosphate, which is used as a negative control to inhibit autophagy.

Advantages of using Premo Autophagy assays
Multiplex inspired — combining the Premo Autophagy Tandem Sensor with the far-red emitting LysoTracker Deep Red allows for a three-color analysis of 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 — Premo Autophagy reagents are non-replicating in mammalian cells, lack observable cytopathic effect, and are suitable for biosafety level (BSL) 1 protocols
Multiple configurations: — Premo Autophagy sensors are available in the following chimeric configurations:
P36235: LC3B-GFP; control LC3B (G120A)-GFP; P36236: LC3B-RFP; control LC3B (G120A)-RFP; P36239: RFP-GFP-LC3B; P36240: GFP-p62; P36241: RFP-p62
For Research Use Only. Not for use in diagnostic procedures.