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This quantity comprises info on examining sphingolipids, sphingolipid shipping and trafficking, and sphingolipid-protein interactions and mobile ambitions. Its significant other quantity 311 offers tools utilized in learning enzymes of sphingolipid biosynthesis and turnover, together with inhibitors of a few of those enzymes, genetic ways, and natural and enzymatic syntheses of sphingolipids and analogs.
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Extra info for Sphingolipid Metabolism and Cell Signaling, Part B
C. Liotta, Anal. Biochem. 17i 373 (1988). 35E. Wilson, E. Wang, R. E. Mullins, D. J. Uhlinger, D. C. Liotta, J. D. Lambeth, and A. H. Merrill, J. Bid. Chem. 263, 9304 (1988). %P. P. Van Veldhoven, W. R. Bishop, and R. M. Bell, Anal. Biochem. 183, 177 (1989). 37H. Ohta, F. Ruan, S. Hakomori, and Y. Igarashi, Anal. Biochem. 222, 489 (1994). 38 K Sato H. Tomura, Y. Igarashi, M. Ui, and F. Okajima, Biochem. Biophys. Rex Commun. 2i0, 32; (1997).  Ceramide Mass Analysis by Normal-Phase High-Performance Liquid Chromatography By ANNEMIEK D.
Rosenthal, and S. Spiegel, Anal. Biochem. 223, 306 (1994). 14B. M. Buehrer and R. M. Bell, J. Biol. Chem. 267, 3154 (1992). 1 ml. The assay tubes are incubated in a shaking water bath at 37” for 30 min, the reaction is stopped by addition of 750 ~1 of methanol, and then brought to 1: 1 methanol : HZ0 (v/v) and loaded onto RP-18 columns and analyzed by HPLC as described in the preceding section. ’ These dual roles make SPP a unique and pivotal lipid in several signal transduction pathways. 2 SPP levels are also regulated via metabolism by an endoplasmic reticulumassociated pyridoxal phosphate-dependent lyase, and also by specific phosphatases, which convert it back to sphingosine.
General Comments 1. Accumulation of excess derivatization reagents and polar lipids on the HPLC column will ultimately alter the chromatographic characteristics and reduce the performance of the column. Routine injection of approximately five sample volumes of 100% 2-propanol at the end of each set of cell samples is therefore recommended to prevent continuous accumulation 22 METHODS FOR ANALYZING SPHINGOLIPIDS 141 of polar material. , Ref. 9) to obtain the neutral (glyco)lipid fraction will considerably lengthen the lifespan of the HPLC column.