AKTA--实验室层析柱装柱技术介绍 PPT课件.ppt

实验室层析柱装柱技术介绍Columnpacking好的柱效是层析成功的关键不同种类的凝胶、ineSepharose4FastFlow5%columnvolsampleColumnpackingmlmAU*ml(volume)%mAUNoRetAreaArea/:N=1600mlmAU*ml(volume)%mAUNoRetAreaArea/:N=3500ColumnpackingN/m=(Ve/Vh)2100/LAs=b/aColumnpacking检测装柱效果一般用<%(30um介质)或<1%(90um介质)柱体积的1%***测柱效和峰形Thelinearflowrateshouldbe30cm/hfor34µmand50µmmediaand20cm/hfor90µmmedia线性流速对体积不同的柱子来说,体积流速不能直接用作比较,一般都换算成线性流速,再作比较: 体积流速(ml/min)x60线性流速(cm/hr)=揪揪揪揪揪揪柱子横切面积(cm2)Columnpacking2019/12/14ColumnpackingItisimportantthatthecolumnisproperlyequilibratedbeforethepackingisevaluated(~2columnvolumes).,,runthecolumnat70%packingpressurefor20hoursandtestitagain(preferablythreetestruns).Notethatpressurespikesmaycausepoorpacking(cracking).Ifthishappens,,必要时重装Columnpacking