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protein_

木虫 (小有名气)

[交流] 【求助/交流】蛋白酶活力的测定方法 已有5人参与

问题是下面的方法中的参比怎么设计,还有更好的测定方法吗?谢谢!
取1mL0.5%酪素,加入0.5mL酶液40℃放置15min,过滤取清液0.5mL加入2.5mL0.55mol/L Na2CO3及0.5mLFollin,40℃显色15min,660nm测吸光值。
空白:取1mL0.5%酪素,加入0.5mL蒸馏水40℃放置15min,过滤取清液0.5mL加入2.5mL0.55mol/L Na2CO3及0.5mLFollin,40℃显色15min(以空白调零)。
酶活力单位定义:上述条件下1min水解1μg酪素产生酪氨酸的酶量即为一个酶活力单位U(绝干)。
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chenyyy

无虫 (正式写手)

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酶先100度水浴失活,或直接加PBS吧,其它相同操作
3楼2011-03-11 13:54:16
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xhjggl

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amisking(金币+1): 鼓励发帖交流。 2011-03-11 20:52:52
可以使用上海生工代理的试剂盒啊!
Protease Assay Kit
蛋白酶活性分析       
Description:
The Protease Assay Kit is designed for the quantitative determination of proteases present in a protein sample, using a dye-labeled protein substrate.The proteases present in the sample of interest will digest the protein substrate and release dye labeled peptides.The absorbance of the dye-labeled peptide is measured at 570nm for determination of protease activity.Chemically stabilized Trypsin (MSG-Trypsin™ is supplied with the kit as a general protease standard; however, other specific protease standards can also be used. MSG-Trypsin™ is an ultra-pure trypsin from bovine pancreas, modified by methylation followed by TPCK treatment and is extremely resistant to autolysis.The kit components are sufficient for 50 assays in a microtiter plate format or 0.5ml assay tubes..application for Determination of protease activity in biological samples, with nanogram detection levels.Store:-15~-20℃
编号        包装        价格        产地
786-028        50 Assays        2112        G Bios
2楼2011-03-11 12:16:04
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4楼2011-05-24 17:54:27
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598286139

铜虫 (正式写手)

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怎么没有加三氯乙酸终止酶反应的那一个啊  对照参比一般加入失活的酶
5楼2013-11-15 18:37:27
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