Plant Lipid Signaling Protocols

Part I: Lipid Analyses

1. Analyzing Plant Signaling Phospholipids through ³²P_i-Labeling and TLC

Teun Munnik & Xavier Zarza

2. Analysis of D3-,4-,5-phosphorylated Phosphoinositides using HPLC

Teun Munnik

3. Mass Measurement of Polyphosphoinositides by Thin Layer- and Gas Chromatography

Mareike Heilmann and Ingo Heilmann

4. Measurement of Inositol (1,4,5) Trisphosphate in Plant Tissues by a Competitive Receptor Binding Assay

Ingo Heilmann and Imara Y. Perera

5. Quantification of Diacylglycerol by Mass Spectrometry

Katharina vom Dorp, Isabel Dombrink and Peter Dörmann

6. Distinguishing Phosphatidic Acid Pools from de novo Synthesis, PLD and DGK

Steven A. Arisz & Teun Munnik

7. Use of Phospholipase A₂ for the Production of Lysophospholipids

Steven A. Arisz & Teun Munnik

8. Analysis and Quantification of Plant Membrane Lipids by Thin-Layer Chromatography and Gas Chromatography

Vera Wewer, Peter Dörmann & Georg Hölzl

9. Lipidomic Analysis of Plant Membrane Lipids by Direct Infusion Tandem Mass Spectrometry

Sunitha Shiva, Hieu Sy Vu, Mary R. Roth, Zhenguo Zhou, Shantan Reddy Marepally, Daya Sagar Nune, Gerald H. Lushington, Mahesh Visvanathan, and Ruth Welti

10. Detection and Quantification of Plant Sphingolipids by LC-MS

Jonathan E. Markham

11. Analysis of Defense Signals in Arabidopsis thaliana Leaves by Ultra Performance Liquid Chromatography/ Tandem Mass Spectrometry: Jasmonates, Salicylic Acid, Abscisic Acid

Nadja Stingl, Markus Krischke, Agnes Fekete and Martin J. Mueller

12. Analysis of Fatty Acid Amide Hydrolase (FAAH) Activity in Plants
Sang-Chul Kim, Lionel Faure, and Kent D. Chapman

13. Ionization Behavior of Polyphosphoinositides Determined via the Preparation of pH Titration Curves using Solid State ³¹P NMR

Zachary T. Graber, and Edgar E. Kooijman

Part II: Enzyme Activities

14. Phosphatidylinositol Synthase Activity from Plant Membrane Fractions and from E. coli-Expressed Recombinant Peptides

Sylvie Collin and Françoise Cochet

15. Phosphatidylinositol 4-kinase and Phosphatidylinositol 4-phosphate 5-kinase Assays

Yang Ju Im, Irena Brglez, Catherine Dieck, Imara Perera and Wendy F. Boss

16. Assaying Inositol and Phosphoinositide Phosphatase Enzymes

Janet L. Donahue, Mustafa Ercetin and Glenda E. Gillaspy

17. Determination of Phospholipase C Activity In Vitro

S.M. Teresa Hernández-Sotomayor and J. Armando Muñoz-Sanchez

18. Assaying Non-Specific Phospholipase C (NPC) Activity

Přemysl Pejchar, Günther F. E. Scherer and Jan Martinec

19. Assaying Different Types of Plant Phospholipase D Activities In Vitro

Kirk L. Pappan and Xuemin Wang

20. Measuring PLD Activity In Vivo

Teun Munnik & Ana M. Laxalt

21. Assaying Plant Phosphatidic Acid Phosphatase Activity

Yuki Nakamura

22. Assay of Phospholipase A Activity

Michael Heinze and Werner Roos

Jonathan E. Markham

Part III: Protein-Lipid Intractions

23. Lipid-Binding Analysis using a Fat Blot Assay

Teun Munnik and Magdalena Wierzchowiecka

24. Liposome-binding Assays to Assess Specificity and Affinity of Phospholipid-Protein Interactions

Magdalena M. Julkowska, Johanna M. Rankenberg and Christa Testerink

25. Lipid affinity beads - From Identifying New Lipid Binding Proteins to Assessing their Binding Properties

Fionn McLoughlin & Christa Testerink

Part IV: Imaging

26. Using Genetically Encoded Fluorescent Reporters to Image Lipid Signaling in Living Plants

Joop E.M. Vermeer and Teun Munnik

27. Imaging Changes in Cytoplasmic Calcium using the Yellow Cameleon 3.6 Biosensor and