Abstract
A simple method for transfer of lipids—including phospholipids, glycolipids, and neutral lipids—from a high performance thin layer chromatography (HPTLC) plate to a polyvinylidene difluoride (PVDF) membrane, TLC-Blot (Far-Eastern Blot), and its biochemical applications are presented. This chapter presents the conventional procedures for separating lipid from tissue samples, cultured cells, and serum and the subsequent development of TLC. Individual lipids separated on an HPTLC plate can be transferred to the PVDF membrane quantitatively and also isolated from the lipid-blotted membrane by a one-step purification procedure. Immunodetection with monoclonal antibodies and treatment with lipid-metabolizing enzymes on the lipid-blotted membrane are possible. The method for identification of individual lipids transferred on the PVDF membrane using matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (TLC-Blot/MALDI-TOF MS) is shown as a functional lipidomics application.
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Acknowledgments
The brain tissues were obtained from The Netherland Brain Bank (NBB) (The Netherlands). All samples were anonymous and were obtained under conditions of informed consent according to the ethical guidelines of the NBB. I deeply appreciate the cooperation of Dr. Rivka Ravid and staff, who were involved in providing the human samples from NBB.
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Taki, T. (2015). TLC-Blot (Far-Eastern Blot) and Its Application to Functional Lipidomics. In: Kurien, B., Scofield, R. (eds) Detection of Blotted Proteins. Methods in Molecular Biology, vol 1314. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2718-0_24
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DOI: https://doi.org/10.1007/978-1-4939-2718-0_24
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2717-3
Online ISBN: 978-1-4939-2718-0
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