? The COBRE for Signal Transduction and Cancer (RR16440) established a Proteomics Facility at West Virginia University. The University donated 900 r2 of renovated lab space. A LC-MS mass spectrometers was loaned to the Proteomics core facility, to initiate this core, while an older Millipore large format 2-D apparatus was donated to the facility. Cobre funds provided for small equipment such as a needle puller, gel equipment and centrifuges. In addition, Cobre funds support personnel, including the core facility director and a technician/mass spectrometry specialist. Subsequently, an equipment grant was funded by the NCRR (S 10-RR16792-01) that provided two dedicated mass spectrometers for the facility, a Maldi-TOF and an LC-MS ion trap mass spectrometer. The cancer center and school of medicine have pledged long term support for the facility, and a steering committee has been established. However, in order to perform proteomic analysis on cancer cells and tissues, the core facility requires additional proteomic-related equipment that enable researchers to identify protein """"""""spots"""""""" of interest from 2-D gels, image and catalog databases of 2-D gels, isolate the individual protein """"""""spots"""""""", digest them and prepare them for sequence analysis. Equipment that perform these functions would help to automate the Proteomics Facility and would seamlessly link with the high throughput capabilities of Maldi and the protein sequencing capabilities of LC-MS ion trap mass spectrometry. In this Cobre supplemental request, we seek bundled major proteomic equipment and software that perform these functions.
Aim 1 : protein purification equipment including HPLC and Millipore water purification.
Aim 2 : a newer 2-D gel apparatus protein separation.
Aim 3 : a scanner, to image 2-D gels, identify protein """"""""spots"""""""" of interest and generate databases, Aim 4: a spot picker, which has associated software that is integrated with the scanner and a robotic arm that can pick individual protein spots from a gel or PVDF.
Aim 5 : a digestion system that permits tryptsinazation and elution of individual proteins.
Aim 6 : computers to provide work stations for data analysis. Collectively, this set of bundled equipment will complete our Proteomics facility. These data will enable our cancer research scientists to identify novel protein targets and proteins that are specifically activated in tumor cell lines and tissues. We hypothesize that this information will enable researchers to assist physicians in customizing cancer treatment to individuals, based on the unique array of signaling proteins found to be activated in an individual tumor biopsy. ? ?
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