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TC-FlAsH™ TC-ReAsH™ II In-Cell Tetracysteine Tag Detection Kit is an expression tag-based fluorescence labeling system for live-cell labeling. The expression construct is created when the Gateway entry clone (containing the gene of interest) is recombined with one of two destination vectors (pcDNA6.2/nTC-Tag-DEST for N-terminal tagging or pcDNA6.2/cTC-Tag-DEST for C-terminal tagging). The expression construct is then used to transform the host strain. The protein can be detected either with green-fluorescent FlAsH-EDT2 reagent or red-fluorescent ReAsH-EDT2 reagent. The tag is very small (6 amino acids) and the protein of interest is fluorescent only when the labeling reagent is added. BAL wash buffer replaces the previously supplied Disperse Blue 3 and EDT-based wash buffer as an olfactorily more agreeable reagent that yields superior signal to noise.
The kit contains FlAsH-EDT2 labeling reagent, ReAsH-EDT2 labeling reagent, BAL wash buffer, pcDNA6.2/cTC-Tag-DEST, pcDNA6.2/nTC-Tag-DEST, and pcDNA6.2/nTC-Tag-p64 Control Plasmid. BAL wash buffer should be stored at 4°C, and the rest of the kit stored at -20°C, protected from light. The kit is stable for 6 months when stored correctly.
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