| Determining Ribosome Translational Status by Ribo-ELISA
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Author:
Date:
2018-01-05
[Abstract] The Ribo-ELISA was originally developed to elucidate the basis for the ribopuromycylation method (RPM)-based detection of ribosome bound nascent chains. The Ribo-ELISA enables characterization of the translational status of ribosomes, and can be applied to the discovery of super-ribosomal complexes with novel ribosome associated macromolecules that are isolated by physical fractionation in sucrose gradients or other methods.
[摘要] Ribo-ELISA最初是为阐明基于核糖核苷酸化方法（RPM）的核糖体结合新生链检测的基础而开发的。 Ribo-ELISA能够表征核糖体的翻译状态，并且可以应用于通过蔗糖梯度中的物理分离或其他方法分离的新型核糖体相关大分子的超核糖体复合物的发现。
【背景】核糖体是由40S和60S亚单位组成的异构结构，当多个核糖体与单个mRNA结合时，它们以单体和多核糖体存在于细胞中。 另外，翻译核糖体可以与调节翻译的多个分子复合物相关联。 核糖体ELISA（Enzyme-Linked ImmunoSorbent Assay）能够通过核糖体相关新生链的体外嘌呤化检测翻译核糖体（David等人，2012）。 在将嘌呤霉素添加至具有结合的新生链的核糖体时，这种化学反应自发进行。 该方法定量测定每个核糖体中出现的新生链的数量，并且可以用于确定单核细胞相对于多核糖体的翻译状态，并鉴定结合其他大分子的核糖体，其改变其在沉淀柱中的沉降速率或迁移。
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| Detection of CLEC5A-JEV Interaction by ELISA
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Author:
Date:
2012-10-05
[Abstract] JEV (Japanese encephalitis virus) belonging to Flaiviridae interacts with CLEC5A (C-type lectin domain family 5, member A), a member of C-type lectin associated with DAP12 signaling protein and expressed on myeloid cell, as the same extent as Dengue virus. This protocol is used to perform and determine the interaction between purified JEV particles with human and murine CLEC5A.Fc fusion proteins by ELISA method. The different strain and batch of purified JEV as well as purity of CLEC5A.Fc fusion protein might affect the result of absorbance. Modifications are most likely needed if users intend to perform ELISA assay.
[摘要] 属于黄病毒科的JEV（ 病毒）与CLEC5A（C型凝集素结构域家族5，成员A） C型凝集素，与DAP12信号传导蛋白相关并且在骨髓细胞上表达，与登革热病毒的程度相同。 该方案用于通过ELISA方法进行和确定纯化的JEV颗粒与人和鼠CLEC5A.Fc融合蛋白之间的相互作用。 不同的菌株和批次的纯化JEV以及CLEC5A.Fc融合蛋白的纯度可能影响吸光度的结果。 如果用户打算进行ELISA测定，则很可能需要修饰。
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