By Karl Maramorosch, Gordon H. Sato
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The cortical layer is stripped from the capsule using a small pair of scissors or curved forceps and the cortex collected into PBS. The cortex is minced thoroughly using two scalpel blades, such that the final tissue pieces are small enough to fit into the bore of a 5-ml pipette. 1% BSA solution and transferred to an Erlenmeyer flask with a sterile amber cork. The flask is shaken on a rotary shaker at 40 rpm for 1 hour at 37°C. The solution is pipetted against the side of the flask three times to free the capillary fragments that have been loosened by the enzyme treatment.
Cell Biol 103, 4 9 - 6 2 . Scholer, H. , and Gruss, P. (1984). Cell 36, 403-411. , and Green, H. (1984). Cell 36, 827-834. Skerrow, C. , and Matoltsy, A. G. (1974). J. Cell Biol 63, 515-523. , Matoltsy, A. , and Matoltsy, M. (1973). J. Biol Chem. 248, 4820-4826. Sporn, M. , Roberts, A. , Wakefield, L. , and Assoian, R. K. (1986). Science 233, 532-534. Steinert, P. M. (1978). J. Mol Biol 123, 4 9 - 7 0 . Steinert, P. , Idler, W. , and Zimmerman, S. B. (1976). J. Mol. Biol. 108, 547-567. Steinert, P.
551 SGECAPNVSVj TVSTSHTSDSGGGSRGGGGGGYGSGGSSYGSGGGSYGSGGGGGGGRGSYGSGGGSYGSGGSSYGSGGGGGGHGSYGSGS . 643 SSGGYRGGSGCGGGGSSGGRGSGGGSSGGSSGGRGSSSGGVKSSGGSSSVKFVSTTYSGVTR SSSQFSSGSQSSRD* V-TSSSRQIRTKVMDVHDGKVVSTHEQVLRTKN . 471 SSSQFSSGSQSSRD V—SSSRQVRTKVVDVHDGKVVFTHEQIVRTKN . nd SSQQASGQSYSSRE VFTSSSSSSSRQTRPILKEQSSSSFSyGySS . R GGGGGRRGGSGGGSYGGSSGGGSYGGSSGGGGSYGGSSGGGGSYGGGSSGCGGRGGG . 513 GGSYGGGRGYGGSSGGGGGGYGGGSSSGGYGGGSSSGGGHGGSSGGSYGGSSSGGGHGGGSSSGGHKSTTTGSVGESSSKGPR NDLSLAKAL . ISNLQQSISDAEQRGENALKDAKNKLNDLEDALQQAKEDLTRLLRDYQELHNTKLALDLEIATYRTLLEGEES.