Description
Antibody to Detect NanoLuc® Luciferase
Anti-NanoLuc® Monoclonal Antibody can be used to detect NanoLuc® Luciferase or NanoLuc® fusion proteins by Western blotting and immunofluorescence. The Anti-NanoLuc® Antibody is a protein A/G affinity-purified mouse monoclonal antibody.
NanoLuc® luciferase is a small (19.1kDa), stable reporter enzyme that generates a bright luminescence signal. The small size and bright signal make NanoLuc® a versatile reporter that is used in many protein analysis applications. NanoLuc® fusion proteins are used for protein stability analysis and in NanoBRET® target engagement and protein interaction applications.
Western Blot Results The figure on the right shows an example Western blot using HEK293 cells transfected with CMV-based expression constructs encoding NanoLuc® fusion proteins. The actual mobility approximated the expected mobility for each fusion protein. For Western blotting, we recommend a concentration of 1µg/ml as a starting point for protocol optimization. Background bands are evident in all lanes, including HEK293 cell lysate, which lacks expression of a NanoLuc® fusion protein, but the NanoLuc® fusion bands are more prominent.
Detection of NanoLuc® fusion proteins expressed in HEK293 cells.
Immunofluorescence Results Shown below are example immunofluorescent images of HeLa cells transfected with a CMV-based construct expressing CDK2-NanoLuc® fusion protein. The CDK2-NanoLuc® fusion protein is stained red, and nuclei are marked blue via Hoechst staining. We recommend a concentration of 1-7.5µg/ml for immunofluorescence optimization. No NanoLuc® staining is observed in HeLa cells transfected with only carrier DNA (mock).
CDK2-NanoLuc® (3µg/ml Anti-NanoLuc®)
NanoLuc®
Hoescht
Merge
Mock (5µg/ml Anti-NanoLuc®)
