Analyzing Protein-Protein Interactions by Co-IP

Protein-protein interactions were analyzed by co-immunoprecipitation (co-IP) as previously described (37 (link)). In brief, anti-E2F1 antibody and protein A/G magnetic beads (Bimake, Houston, TX, USA) were used to immunoprecipitate RB in cell lysates overnight at 4°C. Then the immunoprecipitated beats were collected and washed three times with lysis buffer. The proteins were separated by SDS-PAGE and analyzed by western blotting using anti-RB antibody.

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Yu S., Zhao H., Qin X., Li X., Guo J, & Li W. (2023). Giardia duodenalis-induced G0/G1 intestinal epithelial cell cycle arrest and apoptosis involve activation of endoplasmic reticulum stress in vitro. Frontiers in Immunology, 14, 1127552.

Publication 2023

protein A/G magnetic beads

Anti antibody Buffer Cell Co immunoprecipitation E2f1 Protein a Protein g Proteins Sds page

Corresponding Organization : Northeast Agricultural University

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Variable analysis

independent variables

Anti-E2F1 antibody

dependent variables

Protein-protein interactions between RB and E2F1

control variables

Protein A/G magnetic beads
Lysis buffer
SDS-PAGE
Western blotting using anti-RB antibody

controls

Positive control: Immunoprecipitation of RB using anti-E2F1 antibody
Negative control: Not explicitly mentioned

Annotations

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