Abstract
All living systems depend on metalloproteins. Yet, while tools for the separation and identification of apo-proteins are well developed, those enabling identification and quantitation of individual metalloproteins within complex mixtures are still nascent. Here, we describe the electrophoretic separation of a mixture of carbonic anhydrase, ceruloplasmin, urease, and hemoglobin using native 2D gel electrophoresis and X-ray fluorescence mapping—an approach we have developed to be broadly applicable, not require specialized equipment for sample preparation, and likely to be extensible in the future.
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Acknowledgments
This work was supported by the DOE Office of Science under contract DE-AC02-06CH11357. The authors thank Ken Kemner and Carol Giometti for their support and for initiating efforts in this area at ANL and also thank Stefan Vogt for helpful discussions regarding imaging and for facilitating our efforts at APS.
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Khare, T., Chishti, Y., Finney, L.A. (2012). Electrophoretic Separation and Detection of Metalloproteins by X-Ray Fluorescence Mapping. In: Kurien, B., Scofield, R. (eds) Protein Electrophoresis. Methods in Molecular Biology, vol 869. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-821-4_47
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DOI: https://doi.org/10.1007/978-1-61779-821-4_47
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