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GST Protein Prep.Ver.2 1)Grow 50ml of culture in LB or TB antibiotic o/n at 37℃ shaker. 2)Dilute culture in LB or TB antibiotic 1:10 3)Gr

Low (“Low”)Imidazole Buffer 0.5L 100mM Imidizole 3.4g 5% glycerol 25ml 100% glycerol 50mM Tris-HCl (pH 7.9)50ml 0.5M Tris-HCl (pH 7.9) 0

1)grow 20ml cells O/N37℃ dilute 50X into prewarmed LB,grow to 0.6 OD or about 1hr.Induce w/ 2mM IPTG (238mg/0.5l),grow 3hr,spin 6k GS3 10',f

原理 GST 纯化系统是利用GST （glutathione-S-transferase ）融合蛋白与固定的谷胱甘肽（GSH）通过硫键共价亲和，通过GSH交换洗脱的原理来进行纯化 。1ml树脂大约可结合5-8 mg融合蛋白，并可反复使用数次。试剂u IPTG（异丙基硫代-β-D

Peter Novick Lab,Department of Cell Biology Yale University School of Medicine http://info.med.yale.edu/cellbio/Novick/Second/Protocols/Fus

蛋白质浓缩技术是免疫学中常用的手段，现介绍几种常用的浓缩技术。 （一）透析袋浓缩法 利用透析袋浓缩蛋白质溶液是应用最广的一种。将要浓缩的蛋白溶液放入透析袋（无透析袋可用玻璃纸代替），结扎，把高分子（6 000－12 000）聚合物如聚乙二醇（碳蜡）、聚乙烯吡咯、烷酮等或蔗糖撒

This is a protocol from Engelke et.al.(1990)which was modified by Dr.Baron.The enzyme has been used for RT-PCR,genotyping and cloning. Solu

Solutions 1.1 M Acetic Acid 2.Buffer A Homogenization 10 mM MOPS- 40 ml of 2M stock solution 8 liters 4 mM EDTA- 160ml of 0.2M stock solu

Italics indicate optional steps especially useful for the analysis of untested proteins. GROWTH AND HARVESTING OF BACTERIA Add 100μl ampic

References: Watterson et al.(1980)J Biol Chem 255:962-75. Charbonneau et al.(1979)Biochem Biophys Res Commun 90:1039-47. Extraction of Cal