Mass spectrometric analysis of the electroeluates of fluorescent proteins after preparative electrophoresis in the automated HPGE-1000 apparatus.
Mass spectrometric analysis of the electroeluates of fluorescent proteins after preparative electrophoresis in the automated HPGE-1000 apparatus.
复制标题
在自动化 HPGE-1000 装置中进行制备型电泳后，对荧光蛋白电洗脱液进行质谱分析。
DOI：
10.1002/(sici)1522-2683(19990301)20:3
复制
发表时间：
1999
期刊：
影响因子：
2.9
通讯作者：
Yergey,AL
中科院分区：
文献类型：
--
作者：
Yarmola,E;Chrambach,A;Nguyen,VQ;Yergey,AL
通过电洗脱从凝胶中回收凝胶电泳中的绿色荧光蛋白（GFP）和R-藻红蛋白（PHYCO）条带，所述凝胶电泳在具有周期性荧光扫描的凝胶电泳自动化装置（HPGE-1000装置）上进行。虽然PHYCO在预测的迁移时间后在单一体积的电泳缓冲液中回收，但GFP荧光在相同条件下丢失，并且只能使用多次更换电泳缓冲液来回收。多体积的缓冲液需要合并、浓缩和储存，在这些条件下，少量GFP组分GFP-II人为形成。电洗脱后的PHYCO也显示出在原始制剂中存在的次要组分。在合并、浓缩和储存后转移到质谱仪中的GFP的电泳在质量上与原始制备物不可区分。
Bands of green fluorescent protein (GFP) and R‐phycoerythrin (PHYCO) in gel electrophoresis on the automated apparatus for gel electrophoresis with periodic fluorescence scanning (HPGE), the HPGE‐1000 apparatus, were retrieved from the gel by electroelution. While PHYCO was recovered in a single volume of electroeluate buffer after the predicted migration time, GFP fluorescence was lost under the same conditions and could only be recovered using multiple changes of electroeluate buffer. The multiple volumes of buffer necessitated pooling, concentration, and storage, conditions under which a minor GFP component, GFP‐II, formed artifactually. PHYCO after electroelution also exhibits a minor component present in the original preparation. The electroeluate of GFP, transferred into a mass spectrometer after pooling, concentration and storage, is indistinguishable in mass from the original preparation.