Key Specifications Table
|Key Applications||Detection Methods|
|TFX, IF, ICC||Fluorescent|
|Description||LentiBrite™ GFP-LC3 Control Mutant Lentiviral Biosensor|
|Overview||Read our application note in Nature Methods!
Learn more about the advantages of our LentiBrite Lentiviral Biosensors! Click Here
Biosensors can be used to detect the presence/absence of a particular protein as well as the subcellular location of that protein within the live state of a cell. Fluorescent tags are often desired as a means to visualize the protein of interest within a cell by either fluorescent microscopy or time-lapse video capture. Visualizing live cells without disruption allows researchers to observe cellular conditions in real time.
Lentiviral vector systems are a popular research tool used to introduce gene products into cells. Lentiviral transfection has advantages over non-viral methods such as chemical-based transfection including higher-efficiency transfection of dividing and non-dividing cells, long-term stable expression of the transgene, and low immunogenicity.
EMD Millipore is introducing LentiBrite™ Lentiviral Biosensors, a new suite of pre-packaged lentiviral particles encoding important and foundational proteins of autophagy, apoptosis, and cell structure for visualization under different cell/disease states in live cell and in vitro analysis.
EMD Millipore’s LentiBrite™ GFP-LC3-G120A mutant lentiviral particles serve as a negative control alongside GFP-LC3 wild-type (catalog # 17-10193) for live cell analysis of autophagy.
|Background Information||Autophagy, a degradative pathway that provides recycled nutrients to cells under stress, plays both protective and deleterious roles in many diseases, including cancer, neurodegeneration, and infections. Members of the LC3 family play a key role in the maturation of the autophagosome, the central organelle of autophagy. LC3 precursors, diffusely distributed in the cytosol, are proteolytically processed to form LC3-I. Upon initiation of autophagy, the C-terminal glycine is modified by addition of a phosphatidylethanolamine (PE) to form LC3-II, which translocates rapidly to nascent autophagosomes in a punctate distribution. DNA constructs encoding fluorescent proteins fused to LC3 are widely employed for introduction into cells for monitoring autophagosome formation by fluorescence microscopy.
A mutant form of LC3 with the C-terminal glycine changed to alanine (LC3-G120A) is unable to accept the PE modification and fails to translocate to the autophagosome upon induction of autophagy.
EMD Millipore’s LentiBrite™ GFP-LC3-G120A lentiviral particles serve as a negative control alongside GFP-LC3 wild-type (catalog # 17-10193) for live cell analysis of autophagy.
|Purification Method||PEG precipitation|
|Safety Information according to GHS|
|Product Usage Statements|
|Quality Assurance||Evaluated by transduction of HT-1080 cells and fluorescent imaging performed for assessment of transduction efficiency.|
|Material Size||1 vial (minimum of 3 x 10E8 IFU/mL)|