细胞分离

Column Purification of Demethylated Sphingomyelin

Contributor:SupryaJayadevDate:Mar.11,1991Packingcolumn:1)To20gof100-200meshBio-SilAsilicageladd80mlsofchloroform.2)Placeasmallportionofglasswoolatthebaseofthecolumn.3)Pourgelsolutionintocolumnanduseastirbartopressoutanybubblesintheglasswool.4)Slowlydrainthechloroformwhilestirringtopreventbubblesfrombeingpresentinthepackedgel.5)Add20mlsmorechloroformtorecovertheremainingportionofsilicagelandalsopourintocolumn.6)Leaveasmallportionofchloroformsolutionabovethelevelofpacking.


Columnrun:1)Loadsampledissolvedinchloroformjustabovethepacking.2)Allowsampletopermeatethegelandstarteluting,collectingfractions.3)Elutewith:a)100mlschloroformb)200mlsmethanol-chloroform(1:20)c)200mlsmethanol-chloroform(1:9)d)200mlsmethanol-chloroform(1:5)e)200mlsmethanol-chloroform(1:4)f)800mlsmethanol-chloroform(1:3)g)200mlsmethanol-chloroform(1:3)4)DrydownallfractionsontherotovaporusinghousevacuumandresUSPendinafewmls

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