Nano-Glo® HiBiT Lytic Detection System Technical Manual

Instructions for Use of Product(s)
N3030, N3040, N3050

Literature # TM516

The Nano-Glo® HiBiT Lytic Detection System sensitively quantifies HiBiT-tagged proteins in cell lysates 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 expressed in a cell is determined by adding a lytic 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 in the cell lysate that generates a bright, luminescent enzyme. The amount of luminescence is proportional to the amount of HiBiT-tagged protein in the cell lysate over seven orders of magnitude, with a glow-type luminescent signal that is stable for hours.

Summary of Changes
The following changes were made to the 6/23 revision of this document:
1. Edited Sections 2 and 5.
2. Updated patent statements.
3. Changed font and cover image.
4. Made minor text edits.

Revised 6/23.