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用生化方法制备的呼吸链蛋白系列样品,如HMP,SCR,SDH,Ip,Fp等是生物力能学研究的主要对象。其中微量铁的含量是衡量蛋白纯度的指标之一,亦是FXAFS法测定蛋白中Fe—S结构的前提。此类蛋白制品呈悬浮粘稠液态。SDS可作为良好的溶剂代替繁锁、耗时,易招致损失或污染的酸消化步骤,被应用于直接FAAS标准加入法测定蛋白中微量铁。对于此类蛋白中总血的测定,可直接加入二倍于样品体积的20%SDS溶解,采用标准加入法予以测定,而对含血铁,非血铁的样品,如HMP,SCR则藉生化方法分离之,即用与样品等体积的2N HCl与100℃振荡水浴中保温一小时浸取分离,离心后,分别加入SDS,用于血铁,非血铁的标准加入法测定。
Respiratory chain protein samples prepared by biochemical methods such as HMP, SCR, SDH, Ip, Fp and so on are the main targets of biomechanical studies. The content of trace iron is one of the indexes to measure the purity of protein. It is also the prerequisite for determining the structure of Fe-S in protein by FXAFS. Such protein products were suspended in a viscous liquid. SDS can be used as a good solvent instead of cumbersome, time-consuming, acid-digesting steps that can lead to loss or contamination and is used in the direct FAAS standard addition method for the determination of trace amounts of iron in proteins. For the determination of total blood in such proteins, 20% SDS, twice the volume of the sample, can be dissolved directly and measured by standard addition, whereas biochemical The method of separation, that is, with an equal volume of sample 2N HCl and 100 ℃ shaking water bath for one hour after leaching and separation, centrifugation, were added to SDS, for blood iron, non-ferrous iron standard addition method.