Immunoprecipitation of ELAVL1 complexes

The transfected cells were treated with 20 μM MG132 (Sigma Aldrich) for 6 h, which was followed by cell lysis for immunoprecipitation. The cell lysates were incubated overnight with Protein-A/G MagBeads (Yeasen, Shanghai, China) and anti-ELAVL1 antibody (Cell Signaling Technology) or normal rabbit IgG (Cell Signaling Technology) at 4 °C. The protein complexes were retrieved, washed, and subjected to western blotting with an anti-ubiquitin antibody (Cell Signaling Technology) [23 (link)].

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Xue C., Yang Z., Yang B., Xiong H, & Ye W. (2022). LINC00460 Promotes Cutaneous Squamous Cell Carcinoma Progression Through Stabilizing ELAVL1 Protein. Molecular Biotechnology, 65(8), 1296-1305.

Publication 2022

MG132 Protein-A/G MagBeads anti-ELAVL1 antibody normal rabbit IgG anti-ubiquitin antibody

Anti antibody Cell Immunoprecipitation Mg132 Protein Protein a Rabbit Ubiquitin

Corresponding Organization : Huizhou Central People's Hospital

Other organizations : Guangdong Medical College

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Variable analysis

independent variables

Treatment with 20 μM MG132 for 6 h

dependent variables

Ubiquitination of proteins as measured by western blotting with anti-ubiquitin antibody

control variables

Incubation of cell lysates with Protein-A/G MagBeads and normal rabbit IgG (as a negative control)

controls

Positive control: Incubation of cell lysates with Protein-A/G MagBeads and anti-ELAVL1 antibody
Negative control: Incubation of cell lysates with Protein-A/G MagBeads and normal rabbit IgG

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