Sensitive detection of protein-protein and nucleic acid-protein interactions using a digital imaging system

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Molecular studies of protein-protein interactions have shed invaluable light on a multitude of cellular processes and pathways, ranging from the molecular regulation of intracellular signalling to large-scale understanding of organismal development and life processes. Likewise, studies of how proteins interact with nucleic acids have re-written textbooks and revolutionized our understanding of nucleic acids, both at the level of the nucleic acid itself and at the higher levels of chromatin structure and organization. Mechanistic insights gained from such studies have illuminated our understanding of countless processes and how they regulate life itself. Importantly, work carried out by the Pelka and Kumar groups have used such molecular biology expertise to probe pathways regulating host-pathogen interactions and molecular pathways conferring antibiotic resistance. Our NSERC-funded research focuses on the understanding of how viral proteins reprogram cellular networks during infection and how bacteria disarm antibiotics. This has led to a clearer understanding of not only the mechanisms by which these parasites overtake a host cell, but also have shed important insight into a variety of cellular regulatory networks and the mechanisms by which these networks are controlled, both at the level of the host cell and also within the parasite itself. We depend on an aging Konica SRX 101-A developer with X-ray film for our protein, nucleic acid, and protein-nucleic acid detection needs. It is the key to much of our NSERC-funded research success. Without the ability to detect proteins or nucleic acids we cannot complete the majority of our NSERC Discovery Grant goals. Unfortunately, the system is so old that Konica no longer supports it or manufactures spare parts, as such repairs and service lifetime is very limited while replacement parts are available. In addition, supplies necessary to operate the film developer have become expensive and more difficult to acquire; for example, the developer and fixer chemicals are no longer available in Manitoba and we import them from Ontario at greater cost. Likewise, the costs of film have risen dramatically over the last five years and with the number of western blots we routinely perform, the expense of carrying out these experiments have soared. Combined with frequent down-times due to lack of chemicals, film, or equipment breakdown, this has greatly compromised our ability to perform certain types of assays that rely on sensitive detection of proteins or nucleic acids while harming training of HQP. Ultimately, unrecoverable failure of this system is imminent. Thus, we urgently seek to replace our obsolete film-based Konica SRX 101-A with a new Azure 600 digital imaging system to continue our research and HQP training without fear of irrecoverable equipment failure. Crucially, there is no alternative for us to use and the looming failure of our developer will deprive us of the ability to image blots.