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ral_hope

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[求助] ThT binding assays相关的一句话

Under standard conditions, a stock solution of the
rabbit/human/bovine PrP in 6 M guanidine hydrochloride was diluted to a final
concentration of 10 M and incubated at 37o
C in PBS buffer (pH 7.0) containing 1 M
guanidine hydrochloride and 3 M urea in the absence and presence of different
concentrations of crowding agents with continuous shaking at 220 rpm, and samples
(50 l) were diluted into PBS buffer containing 12.5 M ThT, giving a final volume
of 2.5 ml. The fluorescence of ThT was excited at 450 nm with a slit-width of 7.5 nm
and the emission was measured at 480 nm with a slit-width of 7.5 nm on an LS-55
luminescence spectrometer (PerkinElmer Life Sciences, Shelton, CT).

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ral_hope(金币+5, 翻译EPI+1): 和我想的差不多,但是这个文章还是有问题不能理解,算了,先跳过去了。谢谢啦~ 2011-11-08 08:08:22
在标准状况下,兔/人/猪 PrP 在6M的盐酸胍里的原液被稀释至10mM,保存在37C条件下和(pH=7.0)的PBS缓冲液,其中有1M的盐酸胍和3M的尿素。整个溶液又在无聚集剂和不同浓度的agent里以220转持续震荡。样本(50mL)将在有12.5mM ThT的PBS缓冲液里稀释,总体积为2.5mL。ThT的荧光性在缝隙宽度为7.5nm时在450nm波长为最高峰,在LS-55荧光谱仪上辐射在缝隙宽度为7.5nm,波长为480nm时可以测到。
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