Label IT® Tracker™ Intracellular Nucleic Acid Localization Kits, without TransIT®-LT1 Transfection Reagent

Figure 1. The Label IT® Tracker Kits. The Label IT Tracker labeling reagents are composed of three regions: the label (a fluorophore or biotin) (green), the linker (yellow) which facilitates electrostatic interactions with plasmid DNA and the reactive alkylating group (blue) that covalently attaches the Label IT reagents to any reactive heteroatom within the plasmid. Attachment of the Label IT Reagent to plasmids does not alter their structure or affect downstream gene expression, and as such, the same labeled plasmid can be both visualized after transfection by fluorescence microscopy and employed to express the gene of interest on the plasmid.

Figure 2. Tracking of Plasmid Localization and Expression. COS-7 cells transfected with Label IT Tracker Cy™5 (blue) labeled pEYFP-nuc and TransIT-LT1 Reagent in serum containing media. Images were acquired at 3, 8, and 20 hours post-transfection. The blue signal indicates the cellular localization of the labeled plasmid, the green signal indicates cellular autofluorescence, and the yellow signal is the expression of the nuclear yellow fluorescent protein (YFP) reporter.

Figure 3. Tracking of Plasmid Localization. COS-7 cells were transfected with Label IT Tracker Biotin labeled pEYFP-nuc and TransIT-LT1 Reagent in serum containing media. Image was acquired at 24 hours post-transfection after staining with streptavidin Cy™3 (red) conjugate. The green signal indicates cellular autofluorescence.