Lumit® Immunoassay Cellular Systems Technical Manual
Instructions for Use of Product(s)
W1220, W1201, W1202, W1203, W1231, W1232, W1233, W1331, W1332, W1333
Literature # TM613
The Lumit® Immunoassay Cellular System is a homogeneous bioluminescent assay that measures levels of target proteins in cell lysates when used with the appropriate primary antibody pairs. It combines immunodetection and NanoLuc® Binary Technology (NanoBiT®). NanoBiT® is a structural complementation reporter ideal for protein:protein interaction (PPI) studies. The NanoBiT® System is composed of two subunits: Large BiT (LgBiT; 18kDa) and Small BiT (SmBiT; 11 amino acid peptide), that can be expressed as recombinant fusions or chemically conjugated to target proteins of interest. The LgBiT and SmBiT subunits have been optimized for stability and minimal self-association due to weak affinity (KD:190μM). When two proteins (one tagged with LgBiT, the other tagged with SmBiT) interact, the subunits are brought into proximity to form a functional enzyme that generates luminescence in the presence of its substrate (furimazine).
Summary of Changes
The following changes were made to the 3/24 revision of this document:
1. Lumit® Lysis Buffer II, 10X, a new kit component, was added in Section 2, as well as to flow diagrams and protocols.
2. Lumit trademark was updated to ®.
3. Limited Use Label License (a) and patent statement (b) were updated.
4. Table 3 was added. Figure 7 was modified. New figures 4, 5, 6 and 8 were added.
5. Section 7 was added and remaining sections renumbered.
Revised 3/24.