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文献和实验Co-Expression of human DNA primase p49-p58 subunits
Adapted from W.C.Copeland,Protein Exp.& Purif 9,(1997)1-9 and Schneider,et.al.,JBC 273:34 (1998)21608-21615. Obtained from Fanning lab and revised by LSM,7/23/01 Obtained cell stock of BL21(DE3)cells cotransformed with pET21a-Hp49 (ampicillin
protein, Drosophila Wnt8 (Dwnt8 or WntD), indicating that this protocol is likelyapplicable to all Wnts. Key words: Wnt , Wnt3A , b-catenin , Purification , Blue Sepharose , Immobilized metal affinitychromatography (IMAC) , Gel filtration , Heparin
v468.Chapter 3 分泌型跨膜相关蛋白提纯及Wnt抑制活性
by filtration. 2. Equilibrate heparin resin with 10 column volumes of equilibrationsolution (0.05 M phosphate buffer, pH 7.4, and 0.1M NaCl). For samples derived from as much as 2 L of conditionedmedium, use a 1-mL bed volume and a flow rate of 1mL/min
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