Visualize subcellular structures directly and noninvasively by fluorescence microscopy. With these vectors, you can study cytoskeletal and organelle structure and function in living cells, in real time, and without chemical staining. You can monitor the location of a protein of interest relative to a given subcellular structure by labeling both the protein and the structure with separate fluorescent proteins.
The Subcellular Localization Vectors encode fusions of fluorescent protein variants and localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure. The vectors are available in a variety of organelle- and cytoskeleton-targeted color variants. They are ideal for multiplex labeling experiments.
Photoactivatable mCherry Vectors
PAmCherry is a photoactivatable mutant of mCherry that is non-fluorescent until it is exposed to 350–400 nm light. By selecting which cellular regions to activate, you can track organelles against a dark background. PAmCherry subcellular localization vectors are available that target PAmCherry to the cell membrane, mitochondria, actin, or tubulin, respectively. Observe photoactivated PAmCherry with the same filter sets used for other red fluorescent proteins, such as DsRed variants and mCherry.
Lenti-X Actin Dynamics Monitoring Kit
This kit is designed to monitor the highly dynamic behavior of the actin filament system in live cells. The kit includes lentiviral vectors encoding actin fusions to DD-AcGFP1 (green, destabilized) and mCherry (red), and the DD's stabilizing ligand Shield1. DD-AcGFP1-Actin is continuously targeted for degradation in the cell by the proteasomes, unless the cells are cultured in medium containing the stabilizing ligand Shield1. By contrast, mCherry-Actin (which does not contain the DD) has normal stability upon expression, and is constitutively present in the cell.
Adding and removing Shield1 creates a “pulse-chase” like set of conditions which allow you to monitor polymerization of actin monomers as newly synthesized DD-AcGFP1-Actin that is stabilized by Shield1 (green) is integrated into the existing (red) mCherry-Actin actin filament network.
Autophagy Sensor Vector
pAutophagSENSE is a mammalian expression vector that allows you to monitor the process of autophagy. pAutophagSENSE encodes a fusion of the green fluorescent protein AcGFP1 and the mouse LC3 protein. In cells not undergoing autophagy, the AcGFP1-LC3 fusion protein is evenly distributed in the cytoplasm of a transfected cell. However, if a cell is undergoing autophagy via the formation of autophagosomes, the AcGFP1-LC3 fusion protein is incorporated into autophagosomes. This process can be monitored by following the redistribution of the green fluorescent fusion protein from the cytosol to the forming autophagosomes. The readout is a change from an even cytosolic distribution of green fluorescence in cells to a punctate pattern representing the forming autophagosomes. pAutophagSENSE saves you money compared to other autophagy detection systems, because it is vector-based—not a consumable you have to buy over and over again.