This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project is to provide essential support for HT PDS in terms of sample preparation and handling. The first stage of sample preparation is protein expression, purification, and spin labeling, which is currently accomplished in at least some of the labs of our collaborators with the high efficiency. That is several dozen mutations per week are feasible. At ACERT the samples need to be stored and processed locally as needed, e.g. spin-labeled and freshly reconstituted into liposomes. We shall also develop in this context adequate sample containers compatible with the loading procedures and satisfying the consistency requirements of batch sample processing. With the expected large increase in sample flow and the requirements of fast turn-around, we shall need to increase our sample storage and handling capacities. Also, not all collaborators are able to spin-label their proteins or do it efficiently, therefore those tasks are addressed at ACERT, since we have the expertise and also can assay spin labeling efficiency accurately and expediently using CW and pulse ESR. The success of this project thus heavily relies on the development of suitable facility detailed in subproject 0269.
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