NanoBRET® TE Intracellular RAS Assay Technical Manual
Instructions for Use of Product(s)
N8010, N8012, N8013
Literature # TM717
NanoBRET® Target Engagement (TE) Assays measure compound binding to select target proteins within intact cells and have been applied successfully to study multiple target classes. NanoBRET® TE Assays quantify target engagement using bioluminescence resonance energy transfer (BRET), which is the transfer of energy between a luminescent donor and fluorescent acceptor that only occurs when the donor and acceptor are in close proximity.
The NanoBRET® TE Intracellular RAS Assay measures compound binding to a selected RAS target protein in the presence of cellular factors that affect target engagement potency. Specifically, the assay can quantify apparent intracellular affinity of a test compound for full-length multimeric RAS proteins inside living cells and can be used to study many RAS variants, such as KRAS, HRAS and related mutants.
Whereas the original NanoBRET® target engagement assays used intact NanoLuc® luciferase fused to a target protein as the energy donor, the NanoBRET® TE Intracellular RAS Assay uses the NanoLuc® Binary Technology (NanoBiT®), a structural complementation system comprised of a large luciferase subunit (LgBiT) and a small complementary peptide (SmBiT(q)).
Summary of Changes:
The following changes were made to the 4/24 revision of this document:
1. In Section 4, the definition of BRETMax control was revised.
2. In Section 4.D, additional details were added to Step 2, and Step 3 and 3.C were revised.
3. NanoBRET was updated to a registered trademark.
Revised 4/24.