By Teun Munnik, Ingo Heilmann
As scientist start to comprehend the complexity of lipid signaling and its roles in plant biology, there's an expanding curiosity of their research. a result of low abundancy and temporary nature of a few of those hydrophobic compounds, this isn't constantly effortless. In Plant Lipid Signaling Protocols, expert researchers within the box element experimental techniques during which plant signaling lipids may be studied. those equipment and strategies comprise research of plant signaling lipids, together with unique protocols to observe quite a few appropriate compounds via precise or non-targeted techniques; to assay proper enzyme actions in organic fabric or utilizing recombinant enzymes; to check for particular binding of signaling lipids to protein companions; or to imagine signaling lipids or lipid-derived signs in residing plant cells. Written within the hugely profitable Methods in Molecular Biology sequence structure, chapters comprise introductions to their respective themes, lists of the required fabrics and reagents, step by step, effortlessly reproducible laboratory protocols, and key pointers on troubleshooting and averting identified pitfalls.
Authoritative and practical,Plant Lipid Signaling Protocols aids plant researchers within the carrying on with to review the jobs of lipid signs.
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Extra info for Plant Lipid Signaling Protocols
7. —plate to ensure comigration with lipids from biological material. 8. Dip plate steadily in the CuSO4 solution to avoid tide marks. An automated immersion device can aid uniform dipping. 9. Standard amounts below 1 μg can be visualized using the CuSO4 solution. If no heating plate is available, plates can also be heated more slowly in an oven. 10. To verify linear migration, the scraped remainder of the plate can also be stained by submerging in CuSO4-solution and subsequent heating. 11. The silica powder should not be inhaled, and therefore, make sure to wear protective gear covering your mouth and nostrils.
Based on the properties of the bovine brain adrenal binding protein, the assay is highly specific for Ins(1,4,5)P3 over other inositol phosphates including Ins(1,3,4)P3, Ins(2,4,5) P3, Ins(1,3,4,5)P4, Ins(1,3,4,5,6)P5, and InsP6. One limitation of the protocol presented here is that the Ins(1,4,5)P3 levels in plant cells may be small compared to the total inositol phosphate pool (10), and measurements from unpurified plant extracts may be affected by interference from other compounds present in the sample.
References 1. Welti R, Li W, Li M, Sang Y, Biesiada H, Zhou HE, Rajashekar CB, Williams TD, Wang X (2002) Profiling membrane lipids in plant stress responses. Role of phospholipase D alpha in freezing-induced lipid changes in Arabidopsis. J Biol Chem 277:31994–32002 2. Welti R, Shah J, Li W, Li M, Chen J, Burke JJ, Fauconnier ML, Chapman K, Chye ML, Wang X (2007) Plant lipidomics: discerning biological function by profiling plant complex lipids using mass spectrometry. Front Biosci 12:2494–2506 3.