As exemplified in this SET7/9 study, peptide substrate and enzyme concentrations in the nanomolar range were sufficient to generate robust and reproducible assay signals using either Lance Ultra or AlphaLISA reagents. Figure 3 shows a scheme of how the signal is produced on both platforms. This represents a significant advantage over other technologies using colorimetric or fluorescence-based detection methods that generally require micromolar substrate concentrations.
More significantly, our Lance Ultra and AlphaLISA reagents allowed the use of mid-nanomolar concentrations of SAM, facilitating the screening of SAM competitive inhibitors.
The Lance Ultra europium-antibodies and AlphaLISA Acceptor beads developed for epigenetic research are available as stand-alone reagents, allowing the user maximum flexibility for assay development with preferred substrate and enzyme source. Due to the simple all-in-one-well format, the small number of steps involved (usually 3 or 4), and the absence of wash steps, Lance Ultra and AlphaLISA assays for epigenetic research are amenable to automation. For all these reasons, they constitute a good choice not only for HTS campaigns (hit finding) but also for studies involving secondary screenings and orthogonal testing (hit-to-lead studies).