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

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

Whether protein samples should be pretreated to remove nonspecific binding proteins in co-immunoprecipitation (CO-IP) is controversial. In this work, nonspecific binding of proteins to agarose beads was found to be greater than that to magnetic beads. The nonspecific binding was increased with the d...

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Veröffentlicht in:BioTechniques. - 1993. - 73(2022), 6 vom: 01. Dez., Seite 289-296
1. Verfasser: Zeng, Xi (VerfasserIn)
Weitere Verfasser: Zeng, Wei-Hong, Zhou, Jun, Liu, Xiao-Min, Huang, Guo, Zhu, Hong, Xiao, Shuai, Zeng, Ying, Cao, Deliang
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2022
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Research Support, Non-U.S. Gov't NP-40 Western blot agarose beads co-immunoprecipitation cytoplasmic and nuclear protein extraction nonspecific binding proteins nuclear proteins osmotic pressure mehr... preclear protein–protein interactions Actins Carrier Proteins Nuclear Proteins Tumor Suppressor Protein p53
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520 |a Whether protein samples should be pretreated to remove nonspecific binding proteins in co-immunoprecipitation (CO-IP) is controversial. In this work, nonspecific binding of proteins to agarose beads was found to be greater than that to magnetic beads. The nonspecific binding was increased with the decrease of ion concentrations but reduced by Nonidet P40. Western blot indicated that p65 and β-actin were present as nonspecifically bound protein to the beads. p53 and β-actin were present in the CO-IP precipitates of nuclear proteins but pretreatment cleared the nonspecifically pulled down p53 and β-actin. These data suggest that magnetic beads are better for CO-IP, but preclearing is necessary to minimize false positive regardless of which bead is used, particularly for nuclear proteins 
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650 4 |a Research Support, Non-U.S. Gov't 
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650 7 |a Tumor Suppressor Protein p53  |2 NLM 
700 1 |a Zeng, Wei-Hong  |e verfasserin  |4 aut 
700 1 |a Zhou, Jun  |e verfasserin  |4 aut 
700 1 |a Liu, Xiao-Min  |e verfasserin  |4 aut 
700 1 |a Huang, Guo  |e verfasserin  |4 aut 
700 1 |a Zhu, Hong  |e verfasserin  |4 aut 
700 1 |a Xiao, Shuai  |e verfasserin  |4 aut 
700 1 |a Zeng, Ying  |e verfasserin  |4 aut 
700 1 |a Cao, Deliang  |e verfasserin  |4 aut 
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