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【求助/交流】有荧光标记的多肽在进行低聚物处理后荧光淬灭?
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实验步骤如下,是参考国外文献的: •Solid A peptide was dissolved in cold hexafluoro-2-propanol (HFIP). •The peptide was incubated at room temperature for at least 1 h to establish monomerization and randomization of structure. •The HFIP was removed by evaporation, and the resulting peptide was stored as a film at -20 or -80 °C. •The resulting film was dissolved in anhydrous DMSO at 5 mM and then diluted into the appropriate concentration and buffer with vortexing. •Next, the solution was aged 24 h at 4-8 °C. The sample was then centrifuged at 14000g for 10 min at 4-8 °C; the soluble oligomers were in the supernatant. •Usually, a small pellet was observed suggesting there was some pelletable material in 24 h aggregations. 我照着做了,但是溶入F12以后就形成了絮状物(红色的,因为标记了荧光)。但是放置冰箱的过程中,红色就不见了,絮状物也不见了。为什么呀?离心完就只剩下一点点颜色很浅的沉淀物了。 大家帮忙出出主意吧,谢谢啦 |
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