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基于分子对接虚拟技术及Western blotting实验考察苍耳亭对肝癌上皮间质转化作用靶点的影响 被引量:4
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作者 吴育 曹刚 +2 位作者 石美琴 杨水英 姜晓燕 《药物评价研究》 CAS 2017年第11期1535-1540,共6页
目的用分子对接技术探讨苍耳亭在上皮间质转化(EMT)过程中的作用靶点,并通过Western Blotting实验检测其对肝癌HepG2细胞相应靶点蛋白表达的影响。方法选取在EMT过程中的关键因子蓬乱蛋白(Dhs)、波形蛋白(Vimentin)、Snail、血管内皮生... 目的用分子对接技术探讨苍耳亭在上皮间质转化(EMT)过程中的作用靶点,并通过Western Blotting实验检测其对肝癌HepG2细胞相应靶点蛋白表达的影响。方法选取在EMT过程中的关键因子蓬乱蛋白(Dhs)、波形蛋白(Vimentin)、Snail、血管内皮生长因子受体3(VEGFR3)为作用靶点,采用分子对接虚拟技术评价苍耳亭与其空间结合能力,并与相应内源性物质烟酰胺腺嘌呤二核苷酸、醋酸离子、黄素腺嘌呤二核苷酸、N-乙酰葡糖胺对比;培养HepG2细胞,给予1、5、20μmol/L浓度的苍耳亭,利用Western Blotting实验检测其对Dhs、Vimentin、Snail、VEGFR3蛋白表达的影响。结果分子对接结果显示,苍耳亭与EMT过程中作用靶点均具有一定亲和力,其中与Dhs、Snail、VEGFR3的亲和力高于内源性物质,与Vimentin的亲和能力不及内源性物质;Western Blotting实验结果显示,苍耳亭显著下调Vimentin、Snail、VEGFR3蛋白的表达,显著上调E-cadherin蛋白表达(P<0.05、0.01、0.001)。结论苍耳亭对肝癌侵袭转移关键因子E-cadherin、Vimentin、Snail、VEGFR3有明显影响,可能是其潜在靶点;分子虚拟对接和Western blotting实验结果具有一定的相似性,能预先提示潜在靶因子。 展开更多
关键词 苍耳亭 上皮间质转化 分子对接 Western blotting 肝癌 侵袭转移 作用靶点
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疯牛病和羊痒病Western blotting检测方法的建立 被引量:3
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作者 王辉暖 赵德明 +6 位作者 宁章勇 杨建民 吴常德 郝俊峰 白玉 王传武 孟丽平 《中国兽医学报》 CAS CSCD 北大核心 2007年第1期66-69,73,共5页
以朊蛋白单抗AH6和碱性磷酸酶标记的马抗鼠酶标二抗建立了疯牛病和羊痒病的Western blotting检测方法。对Western blotting各种反应条件进行摸索,并确定了最佳工作条件,结果表明:匀浆缓冲液为RIPA时的最佳反应条件包括浓缩胶电泳电压为... 以朊蛋白单抗AH6和碱性磷酸酶标记的马抗鼠酶标二抗建立了疯牛病和羊痒病的Western blotting检测方法。对Western blotting各种反应条件进行摸索,并确定了最佳工作条件,结果表明:匀浆缓冲液为RIPA时的最佳反应条件包括浓缩胶电泳电压为恒压90V,分离胶电泳电压为恒压160V,转印的最佳电压和时间为恒压100V1.5h;封闭液为3%BSA时,封闭15min,封闭效果最好;AH6的最佳稀释度为1∶4000,4℃下孵育过夜,马抗鼠二抗的最佳稀释度1∶1000,室温下孵育30min。采用已确立的反应条件对样品进行检测并与Prionics-Check WEST-ERN进口试剂盒的检测结果比较,发现其敏感性为100%,特异性为99.4%,与进口试剂盒(100%,100%)无显著差异,这为国产试剂盒研制提供了条件。 展开更多
关键词 疯牛病 羊痒病 WESTERN blotting
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动物源弯曲杆菌免疫显性抗原鉴定及间接ELISA检测方法的建立
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作者 杨欢 王策 +6 位作者 王玉飞 赵晶 张会强 曹易博 姬钰浩 邓兴梅 张辉 《中国畜牧兽医》 北大核心 2026年第3期1418-1433,共16页
【目的】筛选胎儿弯曲杆菌(Campylobacter fetus)、空肠弯曲杆菌(Campylobacter jejuni)血清免疫显性抗原,初步建立动物弯曲杆菌病血清学ELISA诊断方法。【方法】利用生物信息学分析软件对候选抗原进行分析筛选,用pET-28a(+)原核表达载... 【目的】筛选胎儿弯曲杆菌(Campylobacter fetus)、空肠弯曲杆菌(Campylobacter jejuni)血清免疫显性抗原,初步建立动物弯曲杆菌病血清学ELISA诊断方法。【方法】利用生物信息学分析软件对候选抗原进行分析筛选,用pET-28a(+)原核表达载体构建重组质粒,诱导表达并纯化重组蛋白。通过SDS-PAGE和His标签抗体Western blotting检测蛋白表达情况。制备兔源弯曲杆菌多克隆抗体并将其作为Western blotting一抗验证抗原与多克隆抗体反应原性。建立间接ELISA检测方法,优化抗原包被浓度、封闭液及封闭时间、一抗和二抗稀释度及反应时间等反应条件,确定临界值,评价其特异性、灵敏性、重复性。【结果】利用生物信息学分析软件筛选出胎儿弯曲杆菌抗原SapA、空肠弯曲杆菌抗原FlaA。原核表达纯化后,Western blotting检测SapA蛋白大小为101 ku,FlaA蛋白大小为59 ku。制备的胎儿弯曲杆菌兔多克隆抗体血清效价为1∶64000,空肠弯曲杆菌兔多克隆抗体效价为1∶128000。Western blotting结果表明,抗原SapA、FlaA特异性识别弯曲杆菌多克隆抗体,具有良好的反应原性。以纯化的SapA蛋白作为包被抗原,确定的最优包被浓度为2.5μg/mL,封闭液为5%BSA,封闭时间30 min,一抗、二抗稀释度分别为1∶500和1∶8000,孵育时间分别为30和90 min。以纯化的FlaA蛋白作为包被抗原,确定最优包被浓度为0.4μg/mL,封闭时间1 h,其余条件与前者一致。二者与结核分枝杆菌(MTB)、副结核分枝杆菌(MAP)及布鲁氏菌(Brucella)均无交叉反应,特异性强。SapA间接ELISA方法的灵敏性最低达到1∶6400,FlaA间接ELISA方法的灵敏性达到1∶12800。批内和批间重复性试验变异系数均<10%,证明其重复性好。【结论】本研究成功鉴定了胎儿弯曲杆菌免疫显性抗原SapA、空肠弯曲杆菌免疫显性抗原FlaA,建立了动物弯曲杆菌病血清学间接ELISA诊断方法,为弯曲杆菌病防控提供了有效的诊断抗原及快速诊断方法。 展开更多
关键词 弯曲杆菌 免疫显性抗原 原核表达 Western blotting 间接ELISA
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拟南芥DBB1a(double B-box 1a)蛋白的表达、纯化及Western blotting检测 被引量:2
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作者 屠小菊 汪启明 +5 位作者 李秀山 邓克勤 唐东英 罗泽宇 赵小英 刘选明 《激光生物学报》 CAS CSCD 2010年第2期224-228,共5页
PCR扩增拟南芥(Arabidopsis thaliana)DBB1a cDNA的保守区段(GenBank登录号:AT2G21320),转化到冷诱导表达载体pCold TF上,构建pCold-DBB1a重组质粒,转化大肠杆菌DH5a。15℃下IPTG诱导表达融合蛋白,并通过SDS-PAGE检测。证实目的蛋白以... PCR扩增拟南芥(Arabidopsis thaliana)DBB1a cDNA的保守区段(GenBank登录号:AT2G21320),转化到冷诱导表达载体pCold TF上,构建pCold-DBB1a重组质粒,转化大肠杆菌DH5a。15℃下IPTG诱导表达融合蛋白,并通过SDS-PAGE检测。证实目的蛋白以可溶形式在约20 kD处高效表达,与预期蛋白大小相吻合。表达蛋白经Ni琼脂糖凝胶亲和层析纯化,SDS-PAGE及Western blotting检测证实纯化后获得高纯度融合蛋白,这为进一步研究DBBl a功能奠定了基础。 展开更多
关键词 DBBla 原核表达 蛋白纯化 WESTERN blotting
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反向Northern Blotting对小梅山猪下丘脑发育差异表达ESTs的鉴定 被引量:1
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作者 周春宝 潘孝青 +1 位作者 李霖 丁家桐 《江苏农业科学》 CSCD 北大核心 2009年第3期22-24,共3页
用mRNA差异显示技术从小梅山猪下丘脑不同发育阶段,分离并筛选出3个差异表达ESTs,分别命名为MS01、MS02、MS03。利用地高辛DNA标记与检测技术反向Northern斑点杂交,对分离出来的ESTs进行鉴定结果理想。此法避免了同位素放射性污染,操作... 用mRNA差异显示技术从小梅山猪下丘脑不同发育阶段,分离并筛选出3个差异表达ESTs,分别命名为MS01、MS02、MS03。利用地高辛DNA标记与检测技术反向Northern斑点杂交,对分离出来的ESTs进行鉴定结果理想。此法避免了同位素放射性污染,操作简单,可防止DDRT-PCR过程中假阳性的出现,提高了试验准确性,是一种鉴定差异显示ESTs简单而有效的方法。 展开更多
关键词 小梅山猪 反向Northern blotting 下丘脑 ESTS 鉴定
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医学检验专业开设Western Blotting综合性实验初探 被引量:3
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作者 何秋璟 张春龙 侯敢 《卫生职业教育》 2007年第21期91-92,共2页
分子生物学是当前生命科学发展的主流,其理论与技术已广泛应用于各个领域。Western Blottlng作为分子生物学的常规技术,在医学领域具有不可忽视的地位。因此,在医学检验专业开设Western Blotting实验课程,已成为一项非常必要的实验... 分子生物学是当前生命科学发展的主流,其理论与技术已广泛应用于各个领域。Western Blottlng作为分子生物学的常规技术,在医学领域具有不可忽视的地位。因此,在医学检验专业开设Western Blotting实验课程,已成为一项非常必要的实验教学内容。其一,Western Blotting实验是分子生物学与免疫学的有效结合体,有助于学生对前沿的、跨学科知识加深了解; 展开更多
关键词 医学检验专 WESTERN blotting 综合性实验
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原发性肝细胞癌中NSUN6的表达及其临床意义
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作者 叶娟 何建忠 +1 位作者 唐瑶 余继海 《临床与实验病理学杂志》 北大核心 2026年第2期239-243,共5页
目的探讨NSUN6蛋白和mRNA在原发性肝细胞癌(hepatocellular carcinoma,HCC)中的表达及其在HCC进展及预后中的作用和意义。方法使用在线生物信息学分析工具分析NSUN6在HCC患者中的表达,并研究其与HCC患者预后的关系。采用免疫组化法检测9... 目的探讨NSUN6蛋白和mRNA在原发性肝细胞癌(hepatocellular carcinoma,HCC)中的表达及其在HCC进展及预后中的作用和意义。方法使用在线生物信息学分析工具分析NSUN6在HCC患者中的表达,并研究其与HCC患者预后的关系。采用免疫组化法检测91例HCC和43例癌旁组织中NSUN6表达,分析其与HCC临床病理特征的关系。使用Kaplan-Meier生存分析验证NSUN6表达与HCC预后的关系。同时,使用Western bolt和RT-qPCR检测18对HCC癌组织及癌旁组织中NSUN6的表达。结果生物信息学分析示NSUN6在HCC癌组织中明显下调,NSUN6的低表达与更差的HCC预后相关。免疫组化分析表明NSUN6蛋白在HCC癌组织中的低表达率为52.7%(48/91)高于癌旁组织(16.3%,7/43),差异有统计学意义(P<0.05);NSUN6的低表达与分化程度、TNM分期、卫星结节、淋巴结转移均有关(P<0.05)。Kaplan-Meier生存分析示,NSUN6蛋白低表达HCC患者的总生存期低于NSUN6蛋白高表达患者(P=0.007)。Western blot和RT-qPCR检测表明,HCC中NSUN6蛋白及其mRNA的表达均低于配对的癌旁组织,差异有统计学意义(P均<0.05)。结论NSUN6的表达下调可能与HCC密切相关,有望成为疾病进展及预后预测的有效标志物及临床治疗药物研究的新靶点。 展开更多
关键词 肝细胞癌 NSUN6 m5C 免疫组织化学 Western blot RT-QPCR
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细胞转印技术(Cell-Blotting)
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作者 张世馥 《解剖学报》 CAS CSCD 北大核心 1996年第3期303-303,共1页
细胞转印技术(Cell-Blotting)(中国协和医科大学基础医学研究所细胞生物室,北京100005张世馥)自1975年SouthernEM首先建立了Southern转印技术用于DNA的研究之后,相继建立了用于RN... 细胞转印技术(Cell-Blotting)(中国协和医科大学基础医学研究所细胞生物室,北京100005张世馥)自1975年SouthernEM首先建立了Southern转印技术用于DNA的研究之后,相继建立了用于RNA研究的Northern转印以及用... 展开更多
关键词 转印技术 blotting 蛋白分子 培养的细胞 聚丙烯酰胺 基因克隆 蛋白带 单克隆抗体 膜蛋白 蛋白酶抑制剂
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Retraction:Long Noncoding RNA PVT1 Promotes Melanoma Progression via Endogenous Sponging miR-26b
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作者 Oncology Research Editorial Office 《Oncology Research》 2026年第3期766-766,共1页
The published article titled“Long Noncoding RNA PVT1 PromotesMelanoma Progression via Endogenous Sponging miR-26b”has been retracted from Oncology Research,Vol.26,No.5,2018,pp.675–681.DOI:10.3727/096504017X14920318... The published article titled“Long Noncoding RNA PVT1 PromotesMelanoma Progression via Endogenous Sponging miR-26b”has been retracted from Oncology Research,Vol.26,No.5,2018,pp.675–681.DOI:10.3727/096504017X14920318811730 URL:https://www.techscience.com/or/v26n5/56680 Following the publication,concerns have been raised about a number of figures in this article.An unexpected area of similarity was identified in terms of the cellular data,where the results from differently performed experiments were intended to have been shown,although the areas immediately surrounding this area featured comparatively different distributions of cells.In addition,the western blots in this article were presented with atypical,unusually shaped and possibly anomalous protein bands in many cases. 展开更多
关键词 cellular datawhere protein bands cellular data PVT long noncoding RNA MELANOMA miR b western blot
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Retraction:Long Noncoding RNA(lncRNA)HOTAIR Affects Tumorigenesis and Metastasis of Non-Small Cell Lung Cancer by Upregulating miR-613
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作者 Oncology Research Editorial Office 《Oncology Research》 2026年第3期767-767,共1页
The published article titled“Long Noncoding RNA(lncRNA)HOTAIR Affects Tumorigenesis andMetastasis of Non-Small Cell Lung Cancer by Upregulating miR-613”has been retracted from Oncology Research,Vol.26,No.5,2018,pp.... The published article titled“Long Noncoding RNA(lncRNA)HOTAIR Affects Tumorigenesis andMetastasis of Non-Small Cell Lung Cancer by Upregulating miR-613”has been retracted from Oncology Research,Vol.26,No.5,2018,pp.725–734.DOI:10.3727/096504017X15119467381615 URL:https://www.techscience.com/or/v26n5/56685 Following the publication,concerns have been raised about a number of figures in this article.An unexpected area of similarity was identified in terms of the cellular data,where the results from differently performed experiments were intended to have been shown,although the areas immediately surrounding this area featured comparatively different distributions of cells.In addition,the western blots in this article were presented with atypical,unusually shaped and possibly anomalous protein bands in many cases. 展开更多
关键词 non small cell lung cancer cellular datawhere Hotair METASTASIS long noncoding RNA TUMORIGENESIS western blot MIR
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Retraction: MicroRNA-148a Acts as a Tumor Suppressor in Osteosarcoma via Targeting Rho-Associated Coiled-Coil Kinase
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作者 Oncology Research Editorial Office 《Oncology Research》 2026年第1期622-622,共1页
The published article titled“MicroRNA-148a Acts as a Tumor Suppressor in Osteosarcoma via Targeting Rho-Associated Coiled-Coil Kinase”has been retracted from Oncology Research,Vol.25,No.8,2017,pp.1231–1243.DOI:10.3... The published article titled“MicroRNA-148a Acts as a Tumor Suppressor in Osteosarcoma via Targeting Rho-Associated Coiled-Coil Kinase”has been retracted from Oncology Research,Vol.25,No.8,2017,pp.1231–1243.DOI:10.3727/096504017X14850134190255 URL:https://www.techscience.com/or/v25n8/56908 Following the publication,concerns have been raised about a number of figures in this article.An unexpected area of similarity was identified in terms of the cellular data,where the results from differently performed experiments were intended to have been shown,although the areas immediately surrounding this area featured comparatively different distributions of cells.In addition,the western blots in this article were presented with atypical,unusually shaped and possibly anomalous protein bands in many cases. 展开更多
关键词 cellular datawhere tumor suppressor western blots rho associated coiled coil kinase microrna cellular data OSTEOSARCOMA figures
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Corrigendum
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《Neural Regeneration Research》 2026年第3期922-922,共1页
Corrigendum:Epalrestat protects against diabetic peripheral neuropathy by alleviating oxidative stress and inhibiting polyol pathway https://doi.org/10.4103/NRR.NRR-D-25-00562 In the article titled“Epalrestat protect... Corrigendum:Epalrestat protects against diabetic peripheral neuropathy by alleviating oxidative stress and inhibiting polyol pathway https://doi.org/10.4103/NRR.NRR-D-25-00562 In the article titled“Epalrestat protects against diabetic peripheral neuropathy by alleviating oxidative stress and inhibiting polyol pathway,”published on pages 345-351 in Issue 2,Volume 11 of Neural Regeneration Research(Li et al.,2016),the Western blot bands in Figure 2A are incorrect. 展开更多
关键词 western blot bands diabetic peripheral neuropathy alleviating oxidative stress oxidative stress polyol pathway EPALRESTAT
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高内涵分析及Western blotting法在蛋白质核质分布研究中的应用及比较研究 被引量:4
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作者 纪晓方 李丽英 常娜 《首都医科大学学报》 CAS 北大核心 2016年第5期616-620,共5页
目的使用不同方法对蛋白质的核质分布进行检测并比较差异。方法细胞经免疫荧光染色后使用高内涵分析(high content analysis,HCA)对蛋白质的细胞质或细胞核总荧光强度进行分析;分别提取胞质及胞核蛋白,使用Western blotting法对核质组... 目的使用不同方法对蛋白质的核质分布进行检测并比较差异。方法细胞经免疫荧光染色后使用高内涵分析(high content analysis,HCA)对蛋白质的细胞质或细胞核总荧光强度进行分析;分别提取胞质及胞核蛋白,使用Western blotting法对核质组分中的蛋白质含量进行鉴定。结果两种方法均可检测到蛋白质的出核转运;经定量分析,HCA法检测到的HuR蛋白质/核比升高倍数小于Western blotting法。结论使用HCA法检测蛋白质核质定位具有准确、客观、成本低、快捷、多参数等特点,不失为Western blotting的有效补充及替代方法。 展开更多
关键词 高内涵分析 WESTERN blotting 蛋白质核质分布
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肝脏特异性表达载体的构建及Western blotting检测 被引量:1
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作者 张利涛 刘慧琳 曹以诚 《中国畜牧兽医》 CAS 北大核心 2011年第1期100-104,共5页
研究靶向于肝细胞的组织特异性siRNA,实现siRNA基因治疗的组织特异性,可用于特异性治疗乙肝,突破RNA干扰技术在临床应用的一大障碍。用靶向于EGFP的siRNA(以下简称siEGFP)替代靶向于乙肝病毒保守区的siRNA,构建可以在肝细胞中特异性表... 研究靶向于肝细胞的组织特异性siRNA,实现siRNA基因治疗的组织特异性,可用于特异性治疗乙肝,突破RNA干扰技术在临床应用的一大障碍。用靶向于EGFP的siRNA(以下简称siEGFP)替代靶向于乙肝病毒保守区的siRNA,构建可以在肝细胞中特异性表达的载体,用来表达siRNA。将目标载体分别转染肝癌细胞HepG2、乳腺癌细胞MDB-MB-231、人胚肾细胞293,在蛋白质水平上检验siRNA抑制的组织特异性。Western blotting结果证实,siEGFP在肝组织来源的细胞系HepG2中抑制效率明显高于其他两组细胞。构建的载体可以在肝癌细胞系中特异性的表达siRNA,而在其他组织细胞中不表达,实现了组织特异性。进一步将siEGFP替换为抑制HBV表达的siRNA,同样可以实现肝脏特异性的表达。 展开更多
关键词 RNAI 组织特异性siRNA 肝脏特异性 EGFP WESTERN blotting
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Western blotting中低丰度蛋白转膜方法改进 被引量:3
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作者 牟宏宇 罗波 何涛 《泸州医学院学报》 2014年第5期472-475,共4页
目的:改进Western blotting转膜方法,提高低丰度蛋白转膜效率。方法:在Western blotting转膜过程中,采用不同孔径的PVDF膜、不同的转膜时间及不同的甲醇含量进行转膜。转膜后,考马斯亮蓝染色观察PAGE胶上蛋白残留情况,ECL发光法检测FGFR... 目的:改进Western blotting转膜方法,提高低丰度蛋白转膜效率。方法:在Western blotting转膜过程中,采用不同孔径的PVDF膜、不同的转膜时间及不同的甲醇含量进行转膜。转膜后,考马斯亮蓝染色观察PAGE胶上蛋白残留情况,ECL发光法检测FGFR1曝光强度。结果:通过改变膜孔径大小和转膜时间所得实验结果可见,孔径为0.22μm的PVDF膜转移2h的转膜效率更高。通过改变转膜液中甲醇含量所得实验结果可见,甲醇含量为10%转膜效率更高。结论:本实验对Western blotting转膜过程中低丰度蛋白的转膜方法进行了成功改进,改进后能提高蛋白显影强度,具有广泛的应用前景,是一种方便可行的操作方法。 展开更多
关键词 WESTERN blotting 低丰度蛋白质 PVDF膜 甲醇
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A Comparison Between Northern Blotting and Quantitative Real-Time PCR as a Means of Detecting the Nutritional Regulation of Genes Expressed in Roots of Arabidopsis thaliana 被引量:4
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作者 GAN Yin-bo ZHOU Zhong-jing +2 位作者 AN Li-jun BAO Sheng-jie Brian G Forde 《Agricultural Sciences in China》 CAS CSCD 2011年第3期335-342,共8页
Quantitative real-time PCR (qRT-PCR) has become a routine and robust technique for measuring the expression of genes of interest, validating microarray experiments and monitoring biomarkers. However, concerns have b... Quantitative real-time PCR (qRT-PCR) has become a routine and robust technique for measuring the expression of genes of interest, validating microarray experiments and monitoring biomarkers. However, concerns have been raised over the accuracy of qRT-PCR in China as well as in the rest of the world. We have previously used qRT-PCR to study the response of ANR1 and other root-expressed MADS-box genes to fluctuations in the supply of nitrate, phosphate and sulphate under hydroponic growth conditions. In this study, we have used both Northern blotting and qRT-PCR analyses to confirm the nutritional regulation of MADS-box genes in Arabidopsis thaliana and test whether both technologies produce the same results. The information obtained indicated that the qRT-PCR results are consistent with those obtained by Northern blotting hybridization for all the tested root-expressed MADS-box genes, in response to different nitrate, phosphate and sulphate growth conditions. Furthermore, our novel results showed that the expressions of AGL12, AGL18, and AGL19 were all down regulated in response to S and P re-supply in both qRT-PCR and Northern blotting analyses. 展开更多
关键词 Arabidopsis thaliana MADS-BOX nutrient regulation Northern blotting quantitative real-time PCR
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Generation and characterization of an anti-GP73 monoclonal antibody for immunoblotting and sandwich ELISA 被引量:4
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作者 Aixia Zhang Brian Cao 《The Journal of Biomedical Research》 CAS 2012年第6期467-473,共7页
Recently, serum Golgi protein 73 (GP73) levels have been found to be elevated in patients with hepatocellu- lar carcinoma (HCC), and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patien... Recently, serum Golgi protein 73 (GP73) levels have been found to be elevated in patients with hepatocellu- lar carcinoma (HCC), and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patients remain controversial due to the specificity of the anti-GP73 antibody-based enzyme linked immunosorbent as- say (ELISA). Therefore, an anti-GP73 antibody with high specificity was highly demanded. In the present study, by hybridoma screening, we generated an anti-GP73 monoclonal antibody (mAb) designated as 6A2 using recom- binant GP73 protein produced by prokaryotic expression. The specificity of 6A2 was evaluated by Western blot- ting, immunohistochemistry and immunoprecipitation. The results showed that 6A2 recognized GP73 in both native and denatured forms. In addition, we have developed a sandwich ELISA using 6A2 and GP73 polyclonal antibody generated in New Zealand white rabbits according to standard procedures, and measured the serum GP73 level of patients using this assay. Our results showed that serum GP73 levels of HCC patients were significantly higher than those of healthy controls (P = 0.0036). Furthermore, for the first time, GP73 serum level was found to be elevated in patients with breast cancer compared with healthy controls (P = 0.0172). 展开更多
关键词 GP73 monoclonal antibody Western blotting sandwich ELISA hepatocellular carcinoma
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Observation on Plant Leaf Transection by Gelatin Blotting
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作者 TAN Dahai LI Fuheng +1 位作者 WANG Xiaocen HUANG Fushan 《Journal of Northeast Agricultural University(English Edition)》 CAS 2011年第3期7-11,共5页
Blotting was used to observe cell structures of leaf epidermis cells, and the key method of leaf transaction observation was paraffin section. The concentration, suitable solidification time, melting temperature of ge... Blotting was used to observe cell structures of leaf epidermis cells, and the key method of leaf transaction observation was paraffin section. The concentration, suitable solidification time, melting temperature of gelatin solution and the stain for the gelatin blotting were studied in this research. The results showed that the gelatin blotting could be used to study leaf transaction, it was benefit to make operation easily, and save time, money and so on 展开更多
关键词 gelatin solution blotting leaf transaction OBSERVATION
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In Search of Regulators of <i>LeSPL-CNR</i>by South-Western Blotting and Yeast One-Hybrid Library Screening System
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作者 Xiaohong Wang Bishun Ye +2 位作者 Ying Wang Ting Zhou Tongfei Lai 《American Journal of Plant Sciences》 2018年第5期1037-1050,共14页
LeSPL-CNR is a crucial transcription factor for fruit ripening of Solanum lycopersicum. The cnr (colorless non-ripening) epimutation resulted from hypermethylation in a 286 bp region of LeSPL-CNR promoter inhibits nor... LeSPL-CNR is a crucial transcription factor for fruit ripening of Solanum lycopersicum. The cnr (colorless non-ripening) epimutation resulted from hypermethylation in a 286 bp region of LeSPL-CNR promoter inhibits normal fruit ripening. In present study, potential regulators of LeSPL-CNR, which could bind to the specific 286 bp region, were screened via south-western blotting and yeast one-hybrid (Y1H) library screening system. Results indicated that a total of 13 and 19 candidate proteins were acquired respectively, and both ribulose-1,5-bisphosphate carboxylase/oxygenase and 40S ribosomal protein were identified by two methods. These would provide some information for revealing roles of DNA methylation and the regulatory mechanism for LeSPL-CNR. 展开更多
关键词 Solanum lycopersicum LeSPL-CNR 286 bp Region South-Western blotting Yeast One-Hybrid
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Development of a novel protein multi-blotting device
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作者 Amin M. Hagyousif Voon J. Chong +1 位作者 Hiroki Yokota Stanley Y. P. Chien 《Journal of Biomedical Science and Engineering》 2010年第12期1125-1132,共8页
Blotting is a common technique widely used for molecular analysis in life sciences. The Western blot, in particular, is a process of transferring protein samples from a polyacrylamide gel to a blotting membrane and de... Blotting is a common technique widely used for molecular analysis in life sciences. The Western blot, in particular, is a process of transferring protein samples from a polyacrylamide gel to a blotting membrane and detecting the levels of specific proteins through reactions with primary and secondary antibodies. The state-of-the-art of Western blotting usually generates one blotting membrane per gel. However, multiple copies of blots are useful in many applications. Two blotting copies from a single protein gel, for instance, can be used for identifying a total amount of proteins of interest as well as its specific subpopulation level such as a phosphorylated isoform. To achieve this multi-blotting operation from a single gel, we modified a blotting procedure and developed a novel blotting device. The device consisted of a multi-anode plate and a microcontroller. It was designed to generate a well-controlled electrophoretic voltage profile, which allowed a quasi-uniform transfer of proteins of any size. The prototype device was built and its operation procedure was described. The experimental results clearly supported the notion that the described device was able to achieve multiple blotting from a single gel and reduce time and cost for protein analysis. 展开更多
关键词 WESTERN blotting Protein TRANSFER Multi-blotting PWM
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