Multiplexing Click-iT® Plus OPP, GFP, and phalloidin
CellLight® Talin-GFP, BacMam 2.0 (Cat. No. C10611) was used to transduce HeLa cells. After overnight incubation, the Click-iT® OPP reagent was added and incubated for 30 minutes. The cells were then fixed and permeabilized, and treated with Alexa Fluor® 568 Phalloidin (Cat. No. A12380). The labeling of the nascent protein synthesis was completed using the Click-iT® Plus OPP Alexa Fluor® 647 Protein Synthesis Assay Kit (Cat. No. C10458).
Green—indentifies the binding of the talin-GFP fusion protein to the cytoskeleton.
Red—Alexa Fluor®-Phalloidin, identifies the F-actin staining within the cell
Blue punctuate pattern within the nucleus—nascent protein synthesis detected by application of the Click-iT® Plus OPP Alexa Fluor® 647 Protein Synthesis Assay