This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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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