Removal of nonspecific binding proteins is required in co-immunoprecipitation with nuclear proteins

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成果类型：

期刊论文

作者：

Zeng, Xi;Zeng, Wei-Hong;Zhou, Jun;Liu, Xiao-Min;Huang, Guo;...

通讯作者：

Cao, D.;Zeng, Y.

作者机构：

[Liu, Xiao-Min; Zhou, Jun; Zhu, Hong; Zeng, Ying; Xiao, Shuai; Huang, Guo; Cao, Deliang; Zeng, Xi; Zeng, Wei-Hong] Univ South China, Canc Res Inst, Hengyang Med Sch, Hunan Prov Key Lab Tumor Cellular & Mol Pathol, Hengyang 421001, Hunan, Peoples R China.

[Zeng, Ying] Univ South China, Sch Nursing, Hengyang Med Sch, Dept Int & Humanist Nursing, Hengyang 421001, Hunan, Peoples R China.

[Liu, Xiao-Min; Zhou, Jun; Zhu, Hong; Xiao, Shuai; Huang, Guo; Cao, Deliang; Zeng, Xi; Zeng, Wei-Hong] Univ South China, Affiliated Hosp 1, Hengyang Med Sch, Dept pathol, Hengyang 421001, Hunan, Peoples R China.

通讯机构：

[Ying Zeng; Deliang Cao] H

Hunan Province Key Laboratory of Tumor Cellular & Molecular Pathology, Cancer Research Institute, Hengyang Medical School, University of South China, Hengyang, Hunan, 421001, China<&wdkj&>Department of International & Humanistic Nursing, School of Nursing, Hengyang Medical School, University of South China, Hengyang, Hunan, 421001, China<&wdkj&>Hunan Province Key Laboratory of Tumor Cellular & Molecular Pathology, Cancer Research Institute, Hengyang Medical School, University of South China, Hengyang, Hunan, 421001, China<&wdkj&>Department of pathology, The First Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang, Hunan, 421001, China

语种：

英文

关键词：

NP-40;Western blot;agarose beads;co-immunoprecipitation;cytoplasmic and nuclear protein extraction;nonspecific binding proteins;nuclear proteins;osmotic pressure;preclear;protein–protein interactions

期刊：

BIOTECHNIQUES

ISSN：

0736-6205

年：

2022

卷：

期：

页码：

289-296

DOI：

10.2144/btn-2022-0048

基金类别：

This research was funded by the Education Department of Hunan Province with grant no. 20K110 and the Health Commission of Hunan Province with grant no. 202201043124. This research was also funded by the Hunan Changzhutan National Independent Innovation Zone Special Project with grant no. 2018XK2106. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

机构署名：

本校为第一机构

院系归属：

医学院

护理学院

摘要：

METHOD SUMMARYCo-immunoprecipitation is a common method for investigation of protein-protein interactions. However, according to published protocols, it is unclear whether the protein samples should be pretreated with the beads to remove nonspecific binding proteins. The authors investigated the effects of bead types, ion concentrations, NP-40 and pretreatment steps on nonspecifically bound proteins to minimize false positives. Whether protein samples should be pretreated to remove nonspecific binding proteins in co-immunoprecipitation (CO-IP) is controversial. In this work, nonspecific bindin...

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Removal of nonspecific binding proteins is required in co-immunoprecipitation with nuclear proteins

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