The Nano-Glo® HiBiT Dual-Luciferase® Reporter System (HiBiT NanoDLR™) allows sequential measurement of firefly luciferase (Fluc) and HiBiT-tagged proteins from the same sample in an “add-read-add-read” assay format. Fluc is used as a constitutively expressed control to indicate nonspecific compound effects on cell viability or protein expression, rather than specific changes in the levels of the HiBiT-tagged protein of interest, aiding in the interpretation of protein modulation screens and studies.
The 11-amino acid HiBiT peptide tag can be added to a protein of interest using CRISPR/Cas9 gene editing coupled with ectopic expression of Fluc. Alternatively, use a bicistronic HiBiT entry vector to express both the HiBiT-protein fusion and constitutive Fluc on the same mRNA.