Fractionation of mast cell components for studies of ligand-receptor binding at the plasma membrane.
This communication describes a simple procedure for fractionating mast cells producing plasma membranes and intact granules. Mast cells were purified over a bovine serum albumin density gradient and disrupted under conditions in which no histamine was released. Iodinated immunoglobulin E (IgE) bound to the cells served as a marker for the plasma membrane fraction. Employing a discontinuous sucrose gradient the plasma membrane and granule fractions were separated. The specific activity of the IgE binding to the isolated plasma membrane fractions was 10-fold higher compared with that of the IgE binding to intact cells.
D000818:Animals; D002458:Cell Fractionation; D002462:Cell Membrane / Q000032:analysis*; D002499:Centrifugation, Density Gradient; D007073:Immunoglobulin E / Q000378:metabolism*; D008297:Male; D008407:Mast Cells / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D017455:Receptors, IgE; D011971:Receptors, Immunologic / Q000302:isolation & purification / Q000378:metabolism*; D014465:Ultrasonics
D017455:Receptors, IgE; D011971:Receptors, Immunologic; D007073:Immunoglobulin E