Anti-LgBiT Monoclonal Antibody is a protein A/G affinity-purified mouse monoclonal antibody that is used to detect Large BiT (LgBiT) and LgBiT fusion proteins via Western blotting. Weak cross-reactivity with NanoLuc® luciferase is observed.
We recommend a concentration of 1µg/ml as a starting point for protocol optimization.
LgBiT is the large 18kDa subunit of NanoBiT® complementation technology. LgBiT interacts with the low-affinity SmBiT or the high-affinity HiBiT peptides to form a functional luciferase. LgBiT protein fusions are used with SmBiT protein fusions for live cell analysis of protein interactions. Unfused LgBiT can also be expressed in cells to detect HiBiT-tagged proteins without the need for cell lysis.
Western Blot Results
The figure on the right shows an example Western blot using HEK293 cells transfected with CMV-based expression constructs encoding LgBiT fusion proteins. The expected mobility was seen for each of the LgBiT fusion proteins. Low levels of the LgBiT-KRAS fusion were detected even with CMV-driven expression and without dilution using Transfection Carrier DNA.