| 猪凝血酶的分离纯化与鉴定 |
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| 引用本文： | 徐长法 刘德富. 猪凝血酶的分离纯化与鉴定[J]. 北京联合大学学报(自然科学版), 1994, 8(1): 44-49 |
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| 作者姓名： | 徐长法 刘德富 |
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| 作者单位： | 北京大学生物学系，北京联合大学文理学院 |
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| 摘 要： | 本文报导以猪血为材料，制备凝血酶，血浆在低离子强度下经等电沉淀得优球蛋白，从该沉淀中采用稀盐溶液提取，氢氧化镁吸附和二氧化碳解吸等步骤获得凝血酶原粗品。后者经脑提取液激活，再经乙醇分级、ＤＥＡＥ－纤维素与磷酸纤维素离子交换层析和ＳｅｐｈａｄｅｘＧ－１００凝胶过滤，得凝血酶制品，经ＳＤＳ－ＰＡＧＥ分析，制品呈单一条带，分子量约为３９ｋＤ，比活达１６１０ＮＩＨＵ／ｍｇ蛋白，以活化的粗品活性为１００％，活性回收率为４８％，纯化倍数约为１０。
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| 关 键 词： | 猪血 凝血酶 分离 纯化 分子量 |
The Isolation,Purification and Characterization of Thrombin from Porcine Plasma |
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| Xu Changfa,Wang Lanxian,Liu Defu,Ma Xiaolan. The Isolation,Purification and Characterization of Thrombin from Porcine Plasma[J]. Journal of Beijing Union University, 1994, 8(1): 44-49 |
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| Authors: | Xu Changfa Wang Lanxian Liu Defu Ma Xiaolan |
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| Abstract: | The isolation, purification and characterization of pig thrombin were described.Theprocedure includes isoelectric precipitation of euglobulin at low ionic strength,extractiOn of pro-thrombin from the precipitate,adsorption on magnesium hydroxide and subsequent elution with car-bon dioxide at normal atmosphere,activation of prothrombin,ethanol fractionation,chromatographicpurification successively with DEAE-cellulose,phosphocellulose and Sephadex G-100 columns.Thepurified thrombin showed a single band with molecular weight of 39 kD by SDS-PAGE and twobands with isoelectric point values of 5.0 and 5.4 respectively by PAGIF. The specific activity ofthe purified preparation was 1610 NIH U/mg protein.[ |
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| Keywords: | porcine plasma thrombin purification molecular weight isoelectric point. |
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