Description
The BCL-2 TR-FRET Assay Kit is designed to measure the inhibition of BCL-2 (B-cell lymphoma 2) binding to its ligand in a homogeneous 96 or 384 reaction format. This FRET-based assay requires no time-consuming washing steps, making it especially suitable for high throughput screening applications. This kit contains enough recombinant BCL-2, BCL-2 peptide ligand, assay buffer, Anti-His Terbium-Labeled Donor, and Dye-labeled Streptavidin Acceptor for 96 or 384 reactions Figure 1. Illustration of the BCL-2 TR-FRET Assay Kit principle.
A sample containing terbium-labeled donor, dye-labeled acceptor, BCL-2, peptide ligand, and an inhibitor is incubated for 180 minutes. The Donor Terbium-labeled anti-His antibody binds to His-tagged BCL-2. The BCL-2 peptide ligand is labeled with biotin, which allows the dye-labeled streptavidin acceptor to bind to the BCL-2 peptide ligand. The TR-FRET signal is generated by proximity induced upon interaction of BCL-2 with the BCL-2 peptide ligand. Then, the fluorescence intensity is measured using a fluorescence reader capable of TR-FRET measurements. The signal generated is proportional to BCL-2 activity