Evaluating Protein Ubiquitination and Interaction

Immunoprecipitation and ubiquitination evaluation were performed as previously described^{32 (link)}. In brief, cells were treated with MG132 (Abcam) at a final concentration of 20 μm for 4 h before collection. Whole-cell lysates were prepared in RIPA buffer and precipitated using protein A/G beads (Abmart) with β-Catenin antibody (Abmart). Precipitated products were assayed by immunoblot (IB) analysis with anti-Ub antibody (Abcam) or β-Catenin antibody.

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Zhang B., Li Z., Zhang R., Hu Y., Jiang Y., Cao T., Wang J., Gong L., Ji L., Mu H., Yang X., Dai Y., Jiang C., Yin Y, & Zou J. (2019). PKCγ promotes axonal remodeling in the cortico-spinal tract via GSK3β/β-catenin signaling after traumatic brain injury. Scientific Reports, 9, 17078.

Publication 2019

MG132 protein A/G beads β-Catenin antibody anti-Ub antibody

Anti antibody Antibody Buffer Cell Immunoprecipitation Mg132 Protein g Ripa Ubiquitination β catenin

Corresponding Organization :

Other organizations : Nanjing Medical University, Wuxi People's Hospital, Southern University of Science and Technology, Jinan University, Wuxi Institute of Technology

Top 5 similar protocols

Variable analysis

independent variables

Treatment with MG132 (Abcam) at a final concentration of 20 μm for 4 h before collection

dependent variables

Ubiquitination of β-Catenin

control variables

Whole-cell lysates prepared in RIPA buffer
Precipitation using protein A/G beads (Abmart) with β-Catenin antibody (Abmart)

controls

Immunoblot (IB) analysis with anti-Ub antibody (Abcam)
Immunoblot (IB) analysis with β-Catenin antibody

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