UNIT 11.1 Analysis of Inositol Phospholipid Turnover During Lymphocyte Activation
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
How to Cite
Imboden, J. B., Shoback, D. M. and Inokuchi, S. 2001. Analysis of Inositol Phospholipid Turnover During Lymphocyte Activation. Current Protocols in Immunology. 35:11.1:11.1.1–11.1.9.
- Published Online: 1 MAY 2001
- Published Print: FEB 2000
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Receptor-mediated activation of phospholipase C (PLC) leads to the hydrolysis of membrane inositol phospholipids, generating diacylglycerol (DAG) and water-soluble inositol phosphates. This signaling mechanism is used by antigen receptors on T and B cells that have been implicated as mediators of receptor-induced influx of extracellular Ca2+. This unit provides protocols that describe the resolution of InsP by Dowex anion-exchange chromatography. This technique provides a reliable means of separating inositol monophosphate, inositol bisphosphate, and inositol trisphosphate, but does not resolve isomers of these. An Alternate Protocol describes the separation of inositol phosphates by anion-exchange HPLC. A protocol for resolution of inositol phospholipids by thin-layer chromatography (TLC) is also provided.