Protein was collected from cells as previously described (28 (link)). Membranes were blocked in intercept blocking buffer (Li-COR Biosciences). Immunoblotting was performed using mouse anti-FLAG M2 (Sigma-Aldrich), rabbit anti-Actin (Bethyl), or mouse anti-DCAF1 (Proteintech) for 1hr. Blots were incubated with secondary antibodies IRDye 800CW anti-Rabbit and IRDye 680RD anti-Mouse (Li-COR Biosciences) for 1 hr and then visualized using the Li-COR Odyssey M (Li-COR Biosciences).
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Western Blot Analysis of Cellular Proteins
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Corresponding Organization : University of California, Los Angeles
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Variable analysis
independent variables
- Protein collected from cells
- Protein expression levels of FLAG, Actin, and DCAF1
- Membranes were blocked in intercept blocking buffer (Li-COR Biosciences)
- Immunoblotting was performed using mouse anti-FLAG M2 (Sigma-Aldrich), rabbit anti-Actin (Bethyl), or mouse anti-DCAF1 (Proteintech) for 1hr
- Blots were incubated with secondary antibodies IRDye 800CW anti-Rabbit and IRDye 680RD anti-Mouse (Li-COR Biosciences) for 1 hr
- Visualization using the Li-COR Odyssey M (Li-COR Biosciences)
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