Western blotting was performed as described previously (Johnson et al. 2018 (link)). When reprobing blots, HRP-conjugated secondary antibodies were either inactivated by incubation with sodium azide in 5% skim milk/TBST or stripped with Restore Stripping Buffer (ThermoFisher). The following antibodies were used in this study: anti-RACK1 (Cell Signaling, ref. 4716S, 1:1000), anti-RACK1 (Santa Cruz, ref. 17754, 1:500), RPL5 (Genetex, ref. 101821, 1:1000), uS10/RPS20 (abcam, ref. 133776, 1:1000), eS10/RPS10 (Genetex, ref. 101836, 1:1000), uS5/RPS2 (Santa Cruz, ref. 130399, 1:500), and uS3/RPS3 (Bethyl, ref. 303-840A, 1:1000).
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Comprehensive Western Blotting Protocol
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Other organizations : Stanford University
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Variable analysis
independent variables
- Reprobing blots with either HRP-conjugated secondary antibodies inactivated by incubation with sodium azide in 5% skim milk/TBST or stripped with Restore Stripping Buffer (ThermoFisher)
- Protein expression levels of RACK1, RPL5, uS10/RPS20, eS10/RPS10, uS5/RPS2, and uS3/RPS3
- Western blotting was performed as described previously (Johnson et al. 2018)
- Positive controls: Not explicitly mentioned
- Negative controls: Not explicitly mentioned
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