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[求助] α-淀粉酶测定求助

在测定昆虫α-淀粉酶活力的试验中遇到了问题,将提取的酶液稀释不同浓度后加入底物反应,测定出的吸光度没有差异;一开始怀疑是酶量太足将底物反应完全了,固又进行了稀释,稀释到10000倍发现还是同样的问题,吸光度没有差异;然后又测定了一组空白管,不加酶提取液发现同样出现了与上述加入酶液的处理相同的吸光度。本试验底物为1%可溶性淀粉,显色液为3,5-二硝基水杨酸,提取缓冲液为20mmol/L的磷酸钠缓冲液(PH 6.9),采用1mg/ml的麦芽糖标准液测定的标准曲线,相关性很好。究竟问题出在哪里,求大神指导?谢谢!
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