Nano-Glo® HiBiT Extracellular Detection System Technical Manual
Literature # TM523
The Nano-Glo® HiBiT Extracellular Detection System sensitively quantifies HiBiT-tagged proteins expressed in the plasma membrane or secreted into the extracellular medium using a simple add-mix-read assay protocol. HiBiT is an 11-amino-acid peptide tag that is fused to the N or C terminus of the protein of interest or inserted into an accessible location within the protein structure. The amount of a HiBiT-tagged protein present on the cell surface or secreted is determined by adding a nonlytic detection reagent containing the substrate furimazine and Large BiT (LgBiT), the large subunit used in NanoLuc Binary Technology (NanoBiT®). HiBiT binds tightly to LgBiT (KD = 0.7nM), forming a complex that generates a bright, luminescent enzyme. The amount of luminescence generated after adding the Nano-Glo® HiBiT Extracellular Reagent is proportional to the amount of HiBiT-tagged protein accessible to the extracellular medium over seven orders of magnitude.
Printed in the USA 8/17.