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Lipid raft isolation

JR Judith R. Reinhard
AK Alexander Kriz
MG Milos Galic
NA Nico Angliker
MR Mathieu Rajalu
KV Kaspar E. Vogt
MR Markus A. Ruegg
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Lipid raft isolation was performed as described previously25 and modified with minor modifications. After isolation of SMs, sample was adjusted to 45% sucrose. The 45% sucrose sample was overlaid with 12 ml of a linear sucrose gradient (35–5% in 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM CaCl2 and protease inhibitor cocktail (Roche)). Floating lipid raft fractions were isolated by centrifugation at 100,000 g for 20 h at 4 °C.

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