The preparation of a sarcolemmal fraction from evacuated muscle slices.
A novel procedure is described for preparing a plasma membrane fraction from skeletal muscle (e.i., sarcolemma). The procedure entails evacuating the myoplasm from muscle slices as a preliminary step to homogenization and fractionation. The evacuated muscle slices are composed of a stroma-containing sarcolemma, which is then homogenized and fractionated, utilizing a sequence of differential and discontinuous sucrose density gradient centrifugations. On the basis of electron microscopy, selective enzyme markers and alpha-bungarotoxin binding in innervated and denervated muscles, the fraction most enriched with sarcolemma is recovered from the 0.5/0.7 M interface of a discontinuous sucrose gradient.
D000109:Acetylcholine / Q000378:metabolism; D000818:Animals; D002038:Bungarotoxins / Q000378:metabolism; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000648:ultrastructure*; D002499:Centrifugation, Density Gradient; D008297:Male; D008854:Microscopy, Electron; D009132:Muscles / Q000648:ultrastructure*; D051381:Rats; D011950:Receptors, Cholinergic / Q000032:analysis; D012508:Sarcolemma / Q000648:ultrastructure*
D002038:Bungarotoxins; D011950:Receptors, Cholinergic; D000109:Acetylcholine