Convenient and rapid removal of detergent from glycolipids in detergent-resistant membrane microdomains.
Although detergents are often essential in protocols, they are usually incompatible with further biochemical analysis. There are several methods for detergent removal, but the procedures are complicated or suffer from sample loss. Here, we describe a convenient and rapid method for detergent removal from sialic acid-containing glycosphingolipids (gangliosides) and neutral glycolipids in detergent-resistant membrane (DRM) microdomain. It is based on selective detergent extraction, in which the sample is dried on a glass tube, followed by washing with organic solvent. We investigated 18 organic solvents and used high performance thin-layer chromatography (HPTLC) and matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (MALDI-QIT-TOF MS) to confirm that dichloroethane (DCE) was the most suitable solvent and completely removed the nonionic detergent Triton X-100. Furthermore, DCE extraction effectively removed interference caused by other nonionic, zwitterionic, or ionic detergents in MALDI-QIT-TOF MS analysis.
Institute of Glycoscience, Tokai University, Kanagawa, Japan.
Yusuke
Y
Suzuki
Institute of Glycoscience, Tokai University, Kanagawa, Japan. Electronic address: kaba@tokai-u.jp.
Kazuya
K
Kabayama
D041721:3T3-L1 Cells; D000818:Animals; D002855:Chromatography, Thin Layer; D003902:Detergents / Q000737:chemistry* / Q000302:isolation & purification; D005732:Gangliosides / Q000737:chemistry; D006017:Glycolipids / Q000737:chemistry*; D021962:Membrane Microdomains / Q000737:chemistry*; D051379:Mice; D012997:Solvents / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
D003902:Detergents; D005732:Gangliosides; D006017:Glycolipids; D012997:Solvents