Cellular Protein: Protein Interaction Assays Implemented using BacMam
13 May 2014

This seminar will provide an overview of cellular protein: protein interaction methods and the strengths and weaknesses of various assay classes. We have focused on Bioluminescence Resonance Energy Transfer (BRET) assays for detecting and interfering with cellular PPI's and have implemented assays for BH3 family members using BacMam vectors. BacMam vectors afforded an anticipated ease of switching the interacting partners or mutant versions of the partners. In addition, by varying the multiplicity of infection, these vectors provided the ability to increase the sensitivity of the assay by minimally over-expressing the proteins of interest. We will provide examples of the use of this assay and BacMam vectors to measure PPI's using published inhibitors of the interaction of BCL2 and Bad and methods developed for using these vectors for high throughput screening.

Society For Laboratory Automation And Screening