Isolation of lipid rafts from B lymphocytes.
Recent advances in cell biology have provided evidence that the plasma membrane is not a homogeneous lipid bilayer but rather contains within it sphingolipid- and cholesterol-rich membrane microdomains, termed lipid rafts, which serve as platforms for both receptor signaling and trafficking. In B lymphocytes lipid rafts appear to play a key role in the initiation of B-cell antigen receptor (BCR) signaling. Current methods to isolate lipid rafts rely on the relative detergent insolubility of lipid rafts as compared to the nonraft, glycerophospholipid bilayer. Here a method to isolate and characterize lipid rafts from B lymphocytes is described. Particular emphasis is given to the potential artifacts inherent in current procedures that rely on detergents to isolate lipid rafts and alternative technologies that may circumvent these.
Cherukuri
Anu
A
Laboratory of Immunogenetics, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD, USA.
Tzeng
Shiang-Jong
SJ
Gidwani
Arun
A
Sohn
Hae Won
HW
Tolar
Pavel
P
Snyder
Michelle D
MD
Pierce
Susan K
SK
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D002784:Cholesterol / Q000032:analysis*; D003902:Detergents / Q000737:chemistry; D007074:Immunoglobulin G / Q000494:pharmacology; D007075:Immunoglobulin M / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D008223:Lymphoma / Q000276:immunology / Q000378:metabolism; D021962:Membrane Microdomains / Q000737:chemistry* / Q000276:immunology; D051379:Mice; D011947:Receptors, Antigen, B-Cell / Q000032:analysis / Q000276:immunology*; D015398:Signal Transduction; D013107:Sphingolipids / Q000032:analysis*; D014407:Tumor Cells, Cultured
D003902:Detergents; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011947:Receptors, Antigen, B-Cell; D013107:Sphingolipids; D002784:Cholesterol