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作者用大柱制备电泳法从人红细胞解离波中分离提取磷酸葡萄糖变位酶(PGM),并以此为抗原,免疫新西兰家兔,制备得到了兔抗PGM血清.经SDS—PAGE检测,纯PGM为单体,分子量为50,713道尔顿;经琼脂双扩散法检测,兔抗PGM血清效价为1:16,可检出PGM的最低浓度为15.6μg/ml,兔疫电泳表明,该抗血清与PGM2—1型及PGM2—2型人红细胞解离液产生单一的沉淀线,特异性较好。用戊二醛法将该抗血清标记HRP,ELISA测得酶一抗体结合物最适工作浓度为1:128稀释度。 相似文献
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作者应用PGP9.5与抗S100抗体和高压消毒蒸锅抗原复活技术对4例婴幼儿猝死综合征(SIDS)和3例非心源性死亡对照婴儿心传导系统的神经组织总量进行免疫组化研究。结果:在正常婴儿心传导系统,PGP9.5与S100阳性的神经纤维里不均匀分布,以窦房结最多,房室结次之,希氏束最少;4例SIDS心传导系统的神经组织分布同对照组无明显差异。 相似文献
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作者用d-生物素自制N-羟基丁二酰亚胺6-生物素化酰氨已酯(长碳臂活化生物素)。质谱分子离子峰测定分子量为454;红外光谱分析各官能基团与分子结构式吻合。探讨了长碳臂活化生物素与精制马抗银环蛇抗体交联的最适比例。与普通活化生物素标记、辣根过氧化物酶标记的同样抗体相比,稀释滴度分别提高4—8倍。 相似文献
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直接同步扩增5种基因位点 总被引:2,自引:0,他引:2
来用直接同步PCR扩增5种基因位点及反向打点杂交技术,使扩增产物DNA与膜探针条杂交。再通过酶联底物显色反应对LDLR、GYPA、D7S8、Gc和HBGG进行联合分型。该方法特别适用于微量、陈旧或降解的DNA样本,其操作简便、迅速,应用在亲子鉴定案例,亲子关系概率高达99.75%。该技术在法医学、遗传学及临床诊断中具有重要价值,并将改变DNA分型技术难于标准化的局面. 相似文献
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本文以家兔为模型,用酶水解、丙酸酐衍生化处理吸毒后的尿样,用GC/MS同时分析海洛因、吗啡、可待因。 相似文献
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血制和酶型等遗传标记在法医学个人识别与亲权鉴定中有着广泛的应用,目前,一般用免疫学方法和电泳技术对其进行测定,前者需用抗血清,其来源有二:1)天然抗体的分离,如抗-A 抗体可从螺丝和B 型血清中提取.抗-B 抗体可从霉菌和 A 型血清中提取,抗-H 抗体可以从鳝鱼血和荆豆种子浸出液中提取,抗-P 抗体可从山羊、猪等动物的血清中分离得 相似文献
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WTO与GATS的重要区别之一就是它不仅包括货物贸易规则,而且还有投资服务贸易和知识产权等规则。作为第一部管理国际服务贸易的规则,《服务贸易部协定》(GATS)是WTO法律体系的重要组成部分。近年来,我国入世谈判的最主要、最难的议题之一就是服务贸易。从目前的情况看,一方面,在金融服务日益国际化的大趋势下,开放不可避免,更何况开放乃我国基本国策;另一方面,作为一个发展中国家,为保护本国银行业,对外资银行进行一定的限制是必然的,这也是GATS有关规定所允许的,即使是发达国家也有这样或那样的限制。因此… 相似文献
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The study was to eliminate interference from ephedrine in the analysis of methamphetamine. The extraction procedure for methamphetamine was modified to include an oxidation step (2 ml urine specimen was treated with 0.5 ml of 1 mol/L phosphate buffer (pH 6.8) and 0.5 ml 0.3 mol/L sodium periodate). Results showed that ephedrine could be oxidized in the presence of periodate ions into smaller fragments while leaving methamphetamine intact. It is recommended that specimens be treated with sodium periodate prior to extraction in order to eliminate any interference caused by ephedrine. 相似文献
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巴比妥单克隆抗体制备及其免疫学特性鉴定 总被引:1,自引:1,他引:0
目的建立抗巴比妥单克隆抗体杂交瘤细胞株,制备抗巴比妥单克隆抗体,并对其免疫学特性进行鉴定。方法将巴比妥分子环状丙二酰脲环上氨基引入活性羧基,然后通过缩合反应分别与匙孔血蓝蛋白(KLH)和牛血清白蛋白(BSA)偶联,得到完全抗原BAR-KLH和BAR-BSA,并用紫外分光光度计鉴定。用BAR-KLH免疫Balb/C小鼠,利用细胞融合技术建立稳定分泌抗巴比妥单克隆抗体的杂交瘤细胞株;采用体内诱生腹水法制备巴比妥单克隆抗体,饱和硫酸铵法、亲和层析法纯化,并进行免疫学特性鉴定。结果完全抗原偶联成功,其偶联率为30∶1;筛选出1株杂交瘤细胞命名为5C6,其产生的单抗效价为1∶6.4×104,属于IgG1/kappa,抗体亲和力常数为2.27×108L/moL。纯化后的巴比妥单克隆抗体用ELISA法检测其灵敏度为130ng/mL,并与其他7种镇静催眠类药物交叉反应率小于1%。结论本研究制备的杂交瘤细胞株5C6分泌的抗体具有高特异性和灵敏度。 相似文献
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氯胺酮胶体金标记单克隆抗体免疫层析检测板的研制 总被引:7,自引:0,他引:7
目的建立一种简便快速检测氯胺酮的方法。方法将20nm胶体金颗粒标记的抗氯胺酮单克隆抗体,均匀浸在吸水玻璃纤维上,用点膜机将氯胺酮-BSA和纯化后的羊抗鼠IgG多克隆抗体在硝酸纤维薄膜分别划1mm宽的检测线(T线)和质控线(C线),然后依次将吸水玻璃纤维、硝酸纤维薄膜和吸水滤纸粘贴于白色的塑料片上,切成0.5 cm×10 cm大小,制成氯胺酮胶体金标记单克隆抗体免疫层析检测条,并组装成检测板,并检测其特异性和灵敏度。结果氯胺酮胶体金标记单克隆抗体免疫层析检测板在对46种药(毒)物的测试中,仅识别氯胺酮及其主要代谢产物,其准确性为97.6%,检测氯胺酮的阈值为1000ng/m l。结论氯胺酮胶体金标记单克隆抗体免疫层析检测板在5m in内可检出样品中的氯胺酮及其代谢物。 相似文献
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从携带人转铁蛋白单链抗体基因载体转化的大肠杆菌菌株中提取质粒,经电泳分析及筛选、纯化出线状质粒DNA,定量后以载体上人转铁蛋白单链抗体基因插入子两边的已知DNA序列经合成后作为引物进行测序扩增凝胶电泳,将得到的人转铁蛋白单链杭体基因序列重轻链可变区部分同基因库中鼠抗体重轻链进行同源比对,证明人转铁蛋白单链抗体基因确由鼠抗体重轻链可变区基因构成,在此基础上,进一步对基因核苷酸序列的可变区结构特征作推断分析,并推导出其对应的氨基酸序列. 相似文献
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A solid-phase ELISA for Gm typing is described. A mixture of anti-Gm serum (or monoclonal anti-Gm antibody) and test serum was incubated in microtiter wells coated with IgG or its fragments of appropriate Gm type. After washing of the wells, the bound antibody was detected with peroxidase-labeled second antibody. The Glm(3), G3m(16), and G3m(21) antigens could be identified by this technique. Since some of the human anti-Gm sera and anti-Rh0 sera required for the conventional hemagglutination-inhibition method are hard to obtain, the ELISA system using anti-Gm antibodies and no anti-Rh0 sera may serve as an alternative to the conventional method. 相似文献
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Anti-antipyrine antibody was produced in rabbits using 4-azoantipyrine conjugated to bovine serum albumin. The antibody produced against the hapten was examined by the passive haemagglutination test, Ouchterlony analysis and the precipitin test. 相似文献
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D17Z1探针点杂交DNA定量研究 总被引:1,自引:1,他引:0
本研究化学合成了高等灵长类特异性α卫星寡核苷酸片段(D17Z1),经辣根过氧化物酶标记、分子杂交、化学发光法检测,建立了高等灵长类特异性DNA精确定量的方法.制备了DNA浓度梯度标准对照.对人类DNA,猴、猪、牛、羊、鸡、兔、鱼、小鼠等常见动物DNA及JMl09大肠杆菌、入DNA、φ174DNA等微生物DNA进行了定量分析.结果表明,应用该方法对人类DNA定量不受非高等灵长类动物DNA与微生物DNA的影响,可实现组分定量;灵敏度测试,可对0.12ng的人类DNA进行定量,适用于法医DNA检验定量分析. 相似文献
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Simple, rapid methods are described for G3m(21) typing with peroxidase-labeled monoclonal anti-G3m(21) antibody. In G3m(21) typing by ELISA, microtiter wells were coated directly with the test antigen, which was detected with the enzyme-labeled monoclonal antibody. To further simplify the procedure, a dot immunobinding method was developed. The antigen in the test serum applied onto a nitrocellulose membrane was successfully detected with the enzyme-labeled monoclonal antibody. These methods, particularly the dot immunobinding, are suitable for forensic casework because they are rapid and simple and require no technical skill. 相似文献
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Iwadate K Doy M Nishimaki Y Liang F Takatori T Hasekura H 《Forensic science international》2000,110(1):19-28
To examine the use of immunohistochemical staining with antibodies against milk components for detection of aspirated milk on lung sections, eighteen infant death cases were investigated. Immunostaining was performed with anti-human alpha lactalbumin, anti-human IgA, anti-human milk fat globulin 1, and anti-cow whey antibody. Reactivity with each antibody was examined, and semi-quantitative examinations were performed to compare the amount of aspirated milk using anti-human alpha lactalbumin antibody. Materials in the alveoli or bronchioli on lung sections suspected to be aspirated milk showed the most sensitive and clearest reaction with anti-human alpha lactalbumin antibody. Of the eighteen cases, ten cases showed positive reaction with this antibody. The amount of aspirated milk varied widely in each case. In conclusion, immunohistochemical staining with antibodies against human milk components, especially anti-human alpha lactalbumin antibody, can detect small amounts of milk. Using this method, we were able to compare the relative amount of aspirated milk among cases. 相似文献