Immunoprecipitation and Quantitative Western Blotting

Co-immunoprecipitation and Western blot analysis were performed as described previously [20 (link)]. Anti-Flag antibody-conjugated agarose beads (#A2220), anti-HA antibody-conjugated agarose beads (#A2095) and anti-Myc antibody-conjugated agarose beads (#A7470) were purchased from Sigma. Protein G Sepharose (#17–0618–01) was purchased from GE HealthCare Company. The Fuji Film LAS4000 mini-luminescent image analyzer was used to photograph the blots. Image J software (National Institutes of Health) was used to quantify protein levels based on the band density obtained by Western blot analysis.

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Liu X., Deng H., Tang J., Wang Z., Zhu C., Cai X., Rong F., Chen X., Sun X., Jia S., Ouyang G., Li W, & Xiao W. (2022). OTUB1 augments hypoxia signaling via its non-canonical ubiquitination inhibition of HIF-1α during hypoxia adaptation. Cell Death & Disease, 13(6), 560.

Publication 2022

Anti-Flag antibody-conjugated agarose beads anti-HA antibody-conjugated agarose beads anti-Myc antibody-conjugated agarose beads Protein G Sepharose

Anti antibody Co immunoprecipitation Luminescent Protein Protein g Sepharose Western blot analysis

Corresponding Organization :

Other organizations : Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan University, University of Chinese Academy of Sciences

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Variable analysis

independent variables

Anti-Flag antibody-conjugated agarose beads
Anti-HA antibody-conjugated agarose beads
Anti-Myc antibody-conjugated agarose beads

dependent variables

Protein levels based on the band density obtained by Western blot analysis

control variables

Protein G Sepharose

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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