凝胶层析法分离酪蛋白与核黄素

1、2. Gel Chromatography,Separate Casein and Riboflavin by Sephadex G-75,Purpose,basic operation of gel chromatography (凝胶层析,principle of separating riboflavin (核黄素) and casein by Sephadex G-75,welcome to use these PowerPoint templates, New Content design, 10 years experience,1. Conception Gel chromato

2、graphy,inanalytical chemistry, technique for separating chemical substances by exploiting the differences in the rates at which they pass through a bed of a porous, semisolid substance (gel). The method is especially useful for separating enzymes, proteins, peptides, and amino acids from each other

3、and from substances of lowmolecular weight,Gel Chromatography,Gel Filtration (Porath,Flodin,1959,Gel permeation chromatography (GPC,Size-exclusion chromatography (SEC,molecular sieve chromatography (Hjertn,Mosbach,1962,exclusion chromatography (Pedersen, 1962,2. 固定相（凝胶,三维空间网状结构,交联剂越多 交联度越大 网孔结构越紧密,交

4、联剂越少 交联度越小 网孔结构越疏松,Sephadex,Beads are prepared by cross-linking dextran (葡聚糖) with epichlorohydrin (环氧氯丙烷). Widely used in industrial processes. Also useful in desalting (脱盐) and buffer exchange,Sephadex的分级范围 G10 700 G15 1,500 G25 1,000 5,000 G-50 1,50030,000 G-75 3,00070,000 G100 4,000 150,000 G150

5、 5,000 400,000 G200 5,000 800,000,吸水性 *G类的型号 每克干凝胶吸水量（ml）x 10 *如G-75 每克干凝胶吸水量为7. 5 ml,molecular sieve effect (分子筛效应,Ve -Vo Kd = Vi,分配系数Kd,特点： （1）与被分离物质分子的大小和凝胶颗粒孔隙的大小有关 （2）与柱的长短粗细无关,每个溶质分子在流动相和固定相之间有一个特定的分配系数 (Kd,应 用,脱 盐,测分子量,分离纯化,Ve,Albumin,NaCl,Ve,Ve,1) 棉花装填量及松紧度适中； (2) 凝胶不能有气泡和分层现象； (3) 装柱结束后保持凝胶表面平整,1. 装柱,使用滴管，柱床表面以上洗脱液不得流干,Notes