1000ng of vector plasmid DNA encoding pCCL.GFP.T2A.Obsidian luciferase (0.9537kb)
This vector co-expresses a bright green fluorescent protein and a bright thermostable red-shifted bioluminescent ‘infraluciferase’ enzyme termed ‘Obsidian’ and these are expressed in a 1:1 ratio via a self-cleavage sequence (T2A). The green 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 EGFP:
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:
