Complete monitoring of the purification of the plasma membrane from rabbit skeletal muscle.
A fast and reproducible purification procedure for rabbit skeletal muscle plasma membrane is described. Each step was monitored by determination of tetrodotoxin, ouabain and insulin receptors. A ouabain-sensitive K+-stimulated and a Ca2+-dependent phosphatases, probably identical to, respectively the (Na+-K+) and Ca2+-ATPases, were also evaluated. All plasma membrane receptors and the ouabain-sensitive activity accumulated in the lightest fraction separated by sucrose gradient centrifugation (peak at 18% sucrose; purification from crude homogenate, 30-fold).
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D009132:Muscles / Q000378:metabolism / Q000648:ultrastructure*; D010042:Ouabain / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D011817:Rabbits; D011972:Receptor, Insulin / Q000302:isolation & purification; D013779:Tetrodotoxin / Q000378:metabolism
D013779:Tetrodotoxin; D010042:Ouabain; D011972:Receptor, Insulin; D010744:Phosphoric Monoester Hydrolases