LgBiT Expression Vector
The HEK293 LgBiT Cell Line and LgBiT Expression Vector are reagents for CMV-driven intracellular expression of the LgBiT protein, which interacts with the 11-amino-acid HiBiT tag with high affinity to reconstitute the NanoBiT Luciferase enzyme. These intracellular LgBiT expression tools can be paired with HiBiT-tagged proteins expressed either transiently, stably or endogenously via CRISPR-Cas9 insertion to allow endpoint or kinetic real-time monitoring of HiBiT protein levels without the need for cell lysis. Once HiBiT is expressed, NanoBiT enzyme activity is detected with one of the Nano-Glo Live Cell Substrates, which provide options that balance brightness and signal stability. Intracellular LgBiT expression expands the use of the HiBiT tag to live-cell applications such as the measurement of real-time protein dynamics in disease models or following cellular treatment with targeted protein degraders, development of viral infectivity models or assays to monitor intracellular delivery of peptides or small molecules, and use of HiBiT-tagged proteins as the energy donor in NanoBRET interaction assays.