1000ng of vector plasmid DNA encoding pCCL.BFP.T2A.Rhodonite luciferase (0.9537kb)

This vector co-expresses a bright monomeric blue fluorescent protein and a bright thermostable blue-shifted bioluminescent ‘infraluciferase’ enzyme termed ‘Rhodonite’ and these are expressed in a 1:1 ratio via a self-cleavage sequence (T2A). The blue fluorescent protein allows for confirmation of the cassette by fluorescence microscopy and flow cytometry and the bioluminescent enzyme allows sensitive in vitro and in vivo bioluminescence detection and imaging.

Fluorescence microscopy of BFP:

Rhodonite and Obsidian unmixing with D-luciferin:

Rhodonite and Obsidian unmixing with D-infraluciferin methyl ester (obtained from Bioflares):

Light yield of reporters in HEK293T cells with 1mM D-luciferin or D-infraluciferin me ester in phosphate buffered saline (PBS). Bioflares' D-infraluciferin ester gives 6% of the light with D-luciferin with these bright reporters: