The Role of Inositol and the Principles of Labelling, Extraction, and Analysis of Inositides in Mammalian Cells
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Inositides have an important impact on diverse areas of cellular regulation. However, since this area has grown exponentially from the mid 1980s onwards, many workers find themselves relatively new to the field. In this chapter, we establish a broad foundation for the rest of the book by covering some important principles of inositide methodologies. The focus is especially directed to those methods or aspects of methodology not covered in detail in other chapters. This includes the often neglected influence of the inositide precursor, inositol, and important background information relating to the labelling and extraction of inositides from cells and tissues. This introductory section also gives a “birds eye” view of important methods and protocols found within this volume and hopefully acts as a touchstone to assess which of the methodologies described within this book is most appropriate for your particular study(ies) of inositides.
Key wordsmyo-inositol D-myo-inositol 3-phosphate synthase Inositol phosphate Inositol lipid Equilibrium labelling Acid extraction
The authors would like to thank Prof. R.H. Michell and Dr. C.J. Kirk for permission to use unpublished data obtained while C.J.B was a research fellow in their laboratory and Prof. R.F. Irvine for reading and commenting on the manuscript. This work was supported by grants from Karolinska Institutet, Novo Nordisk Foundation, the Swedish Research Council, the Swedish Diabetes Association, EFSD, The Family Erling-Persson Foundation, Berth von Kantzow’s Foundation, and EuroDia (LSHM-CT-2006-518153).
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