Seminar Abstract

“Application of Reverse Phase Protein Arrays to discovering the distribution of important proteins in the transmural proteome of the canine left ventricle”

Reverse Phase Protein Arrays (RPPAs) are a relatively new method utilized in quantitative proteomics. Virtually all RPPA experiments are used to detect the concentration of a target protein present in relatively low abundance among a milieu of many higher abundance proteins extracted from a cell culture or tissue sample. RPPAs are unique in that they immobilize the entire protein extract on a nitrocellulose surface via robotic printing and probe the extract for the target protein with a specific antibody. These arrays will be utilized to measure the transmural distribution of proteins important in the excitation-contraction coupling process of the canine left ventricle. Due to the properties of RPPAs, it is hypothesized that we will be able to measure the distribution of proteins with greater statistical power and spatial resolution than previously achieved. Prior to the use of the arrays in the discovery phase, optimization and validation experiments needed to be carried out in order to ensure the worth of the collected data. In this talk, I will explore RPPAs in detail and share with you some of the results from optimization and validation of the arrays through spike-in protein studies and subsequent computational analysis. Furthermore, I will outline the future experiment which will collect transmural protein distribution data.