Co-immunoprecipitation and semi-quantitative immunoblotting for the analysis of protein-protein interactions.
Co-immunoprecipitation and semi-quantitative immunoblotting for the analysis of protein-protein interactions.
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DOI：
10.1016/j.xpro.2021.100644
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发表时间：
2021-09-17
期刊：
影响因子：
--
通讯作者：
Danuser G
中科院分区：
文献类型：
--
作者：
Burckhardt CJ;Minna JD;Danuser G
来自细胞裂解物的蛋白质复合物的免疫共沉淀（co-IP）被广泛用于研究蛋白质-蛋白质相互作用。然而，建立稳健的co-IP测定通常涉及相当大的优化。此外，co-IP结果经常以非定量的方式呈现。该方案提出了一种优化的co-IP工作流程，其分析基于半定量免疫印迹密度测定法，以提高可靠性和重现性。有关本方案使用和执行的完整详细信息，请参阅。免疫共沉淀法定量细胞裂解物中蛋白质相互作用半定量免疫印迹密度法用于扰动的统计评估优化的工作流程使用标准实验室设备在四天内得出结果细胞裂解物中蛋白质复合物的免疫共沉淀（co-IP）广泛用于研究蛋白质-蛋白质相互作用。然而，建立稳健的co-IP测定通常涉及相当大的优化。此外，co-IP结果经常以非定量的方式呈现。该方案提出了一种优化的co-IP工作流程，其分析基于半定量免疫印迹密度测定法，以提高可靠性和重现性。
Co-immunoprecipitation (co-IP) of protein complexes from cell lysates is widely used to study protein-protein interactions. However, establishing robust co-IP assays often involves considerable optimization. Moreover, co-IP results are frequently presented in non-quantitative ways. This protocol presents an optimized co-IP workflow with an analysis based on semi-quantitative immunoblot densitometry to increase reliability and reproducibility. For complete details on the use and execution of this protocol, please refer to. Co-immunoprecipitation protocol to quantify protein interactions in cell lysates Semi-quantitative immunoblot densitometry for statistical assessment of perturbations The optimized workflow gives results in four days with standard lab equipment Co-immunoprecipitation (co-IP) of protein complexes from cell lysates is widely used to study protein-protein interactions. However, establishing robust co-IP assays often involves considerable optimization. Moreover, co-IP results are frequently presented in non-quantitative ways. This protocol presents an optimized co-IP workflow with an analysis based on semi-quantitative immunoblot densitometry to increase reliability and reproducibility.
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影响因子：
3.3
作者：
Pollard, Daniel A.;Pollard, Thomas D.;Pollard, Katherine S.
通讯作者：
Pollard, Katherine S.
影响因子：
2.9
作者：
Gassmann, Max;Grenacher, Beat;Vogel, Johannes
通讯作者：
Vogel, Johannes
影响因子：
3.7
作者：
Degasperi A;Birtwistle MR;Volinsky N;Rauch J;Kolch W;Kholodenko BN
通讯作者：
Kholodenko BN
DOI：
10.1007/978-1-4939-2425-7_25
发表时间：
2015-01-01
期刊：
PROTEIN-PROTEIN INTERACTIONS: METHODS AND APPLICATIONS, 2ND EDITION
影响因子：
--
作者：
Takahashi, Yoshinori
通讯作者：
Takahashi, Yoshinori
DOI：
10.1007/978-1-4939-7033-9_17
发表时间：
2017-01-01
期刊：
BACTERIAL PROTEIN SECRETION SYSTEMS
影响因子：
--
作者：
Lin, Jer-Sheng;Lai, Erh-Min
通讯作者：
Lai, Erh-Min