纯化柱的选择Superdex200分析型分子筛柱位阀凝胶过滤层析分子筛）Gel Filtration脱盐柱Hitrapn5mlDesatting唐门大了止干至姜糯无典
自强不息 止于至善 纯化柱的选择 Gel Filtration 脱盐柱 Superdex200 分析型分子筛 柱位阀
科命生学分子筛（凝胶过滤层析）格1.Sphericalparticlespacked intoa column.根据分子的大小不同来分离的，大分子由于进入不到填料孔径内，走的路径短所3以先从层析柱中2.Sampleapplied.3.Buffer(mobilephase)andsample4.Largemoleculesleavethecolumn流出来，小分子firstfollowedbysmallermoleculesinmovethroughcolumn.Moleculesdiffuseinandoutofmatrixorderofsize.Moleculeslargerthanthe相反。matrixporespassstraightthroughiDsionot废门大王强子典止干至1
自强不息 止于至善 1. Spherical particles packed into a column. 2. Sample applied. 3. Buffer (mobile phase) and sample move through column. Molecules diffuse in and out of matrix 4. Large molecules leave the column first followed by smaller molecules in order of size. Molecules larger than the matrix pores pass straight through. Diffusion Buffer Diffusion into the pores Diffusion out of the pores Buffer 分子筛（凝胶过滤层析） 根据分子的大小 不同来分离的， 大分子由于进入 不到填料孔径内 ，走的路径短所 以先从层析柱中 流出来，小分子 相反
ForverysmallFor peptides and globularForlargermoleculesmoleculesproteinsorparticles1k100k10k1000k102103104105106107108SuperdexPeptideSuperdex 30prepgradeSuperdercpreegedSuperdex 200 prep grade不同填料有Sephacryl S-100High ResoluticSephacrytS-200HighResotutionSephacryS-3ooHioh Reso特定的分离SephacryfS-400HighResoiutioephacryiS-500HighResolutionSephacryi S-1000S范围Sepharose6FastFlowSepharose 4Fast FiowSepharose CL-28Sepharose CL-4BSepharoseCL-68Superose6Superose12Sephadex G-10Sephadex G-15SephadexG25FS-超圳SephatexG-25MSephadexG-25CF-SephedixG-25SFM-中SephadexG-50FSephadex G-50MC=粗Sephadex G-50 CSephadexG-50SSephadexG-75Sephadex G-75 SFSephadex G-100SephadexG-100SFSephadex G-150唐门大了止干舌真光电
自强不息 止于至善 1k 10k 100k 1000k 不同填料有 特定的分离 范围
Superdex200过柱流速的选择分析型分子筛脱盐柱HTrap5ml根据纯化柱说明书进行Desalting选择5 ml柱：一般5 ml/minColumn dataTable 1. HiTrap Desalting characteristicsMotrixComposite of cross-linked ogorose ond dextronMatrixSephadex G-25 Superfine.Bed dimensions10300310mmcross-linked dextranBedvolumeApproximotely 24 ml1ml柱：一般1ml/minSeparation mechanismAccordingtosizeColumn efficiency, N/M>30 000m13 jmColumn volume5mlAverage ParticleSizepH stobility rongeColumn dimension1.6 ×2.5 cm181regularustVoid volume1.5 mlcleaningTemperotureRecommended sample volume0.1-1.5ml分析型分子筛4 *C to 40 *Coperoting1.3-4.0×applied volumeSompledilution,syringeoperation4°Cto30*CstorogeM.5000Exclusion limitSuperdex75Superdex 2000.5-1 ml/minBeadsize15-70μmExcusion limit, M,Approx.1×10%Approx13×10mglobudlor proteinsMaximumflow rote*15ml/minOptimumseporation.tonge1-10 ml/minRecommendedflowrote*3 00070 00010 000600 000alobulorproteins,MBack pressureat10 ml/min*0.25 bardextrons50030 0001000100 000Maximum backpressure3 bor,0.3 MPoFlow rate (water at room temperaturel)制备型分子筛0.5-10 m/min0.25-0.75 ml/minrecommendedAllcommonlyused buffersChemicalstabilitymoximun1.5ml/min1ml/min2-13pH stobility.short ond long termPressure over columrStorage20% ethanol4ml/min1.,8 MPo, 18 bor, 261 psi1.5 MPo,15 bor, 218 psimaiimum度门大王止干至美招子电
自强不息 止于至善 过柱流速的选择 • 根据纯化柱说明书进行 选择 • 5 ml柱：一般5 ml/min • 1 ml柱：一般1 ml/min • 分析型分子筛 0.5-1 ml/min • 制备型分子筛 4ml/min 脱盐柱 Superdex200 分析型分子筛
上样方式·上样环：0.01~5ml上样环?超级上样环：1ml~150ml上样泵系统泵·上样泵：0.5~1L·系统泵：>50ml废门大王强不息上干至善
自强不息 止于至善 上样方式 •上样环：0.01~5ml •超级上样环: 1ml~150ml •上样泵: 0.5~1L •系统泵: >50ml 系统泵 上样环 上样泵