在制备尖吻蝮蛇毒出血毒素抗血清中不同凝胶染色方法的比较.pdf

韦传宝等摇 在制备尖吻蝮蛇毒出血毒素抗血清中不同凝胶染色方法的比较摇 第12期 ·1793 · doi:10.3969/ j.issn.1000鄄484X.2016.12.015 在制备尖吻蝮蛇毒出血毒素抗血清中不同凝胶染色方法的比较淤 韦传宝摇 曹佳敏摇 (皖西学院生物与制药工程学院,六安 237012) 摇 摇 中图分类号摇 R991摇 摇 文献标志码摇 A摇 摇 文章编号摇 1000鄄484X(2016)12鄄1793鄄04 [摘摇 要]摇 目的:寻找在制备出血毒素抗血清中电泳后染色凝胶的好方法。 方法:按照前人的方法从皖南产尖吻蝮蛇 毒中初步分离3种出血毒素:AaH 玉、AaH 域和AaH 郁并获得3种粗提取的出血毒素,用制备电泳的方法进一步纯化AaH玉、 AaH 域和AaH 郁,对每种出血毒素,配制6块制备电泳板并进行聚丙烯酰胺凝胶电泳,用6种不同的染色方法分别染色,将含 出血毒素的凝胶切下,磨细后直接免疫注射小鼠并获得抗血清,用ELISA检测和中和出血毒素出血活性两种方法检测抗血清 的质量。 结果:不同染色方法制备的抗血清中有效 IgG浓度不同,用无毒型蛋白质快速染色试剂盒染色制备的抗血清有效 IgG浓度最高。 结论:用无毒型蛋白质快速染色试剂盒染色是制备抗血清最好的染色方法。 [关键词]摇 尖吻蝮蛇;出血毒素;抗血清制备;染色法;质量检测 Comparative studies by different dyeing methods on antiserum preparation against hemorrhagins from Agkistrodon acutus venom WEI Chuan鄄Bao,CAO Jia鄄Min郾Biological and Pharmaceutical Engineering College,West Anhui University,Liuan 237012,China [Abstract]摇 Objective:In order to look for a good method for preparation of hemorrhagin antiserum郾Methods:Three kinds of hemorrhagins including AaH 玉,AaH 域,and AaH 郁 were purified from Agkistrodon acutus venom according to predecessors忆s methods and crude AaH 玉,AaH 域and AaH 郁were obtained郾 Preparation electrophoresiswas used to purify AaH 玉,AaH 域and AaH 郁further郾 Asfor an hemorrhagin,six different dyeing methods were used to dye PAGE gel and the gel contained hemorrhagin was obtained respectively郾 Thegroundgelcontainedhemorrhaginwasusedtoimmunemiceanditsantiserumwasobtained郾 Antiserums quality was testedthrough ELISAtest andneutralization of thehemorrhagic activitiesof correspondinghemorrhagin郾 Results:Effective IgG concentration in different antiserum was different and effective IgG made through non toxic type protein fast stain reagent kit was higher than others郾 Conclusion:Nontoxictypeproteinfast stainreagent kit isthebest dyeing method amongthe six dyeing methods郾 [Key words]摇 Agkistrodon acutus;Hemorrhagin;Antiserum preparation;Dyeing;Quality test 摇 摇 尖吻蝮蛇(Agkist