Immunoblotting and qPCR were done exactly as described previously (Jana et al. 2017 (link)). We used anti-Pus10 (HPA049582, Sigma) antibody for the immunoblots shown in Figure 3 C and Supplemental Figure S5B . We also used another commercial anti-Pus10 (HPA044736, Sigma) antibody, but it showed too many nonspecific bands in both nuclear and cytoplasmic extracts and was not useful. Anti-HisTag antibody (2365S, Cell Signaling) was used to check for overexpressed Pus10.
Protocol detail
Immunoblotting and qPCR analysis of Pus10
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Anti antibody
Antibody
Cytoplasmic
Immunoblots
Corresponding Organization :
Other organizations : Southern Illinois University Carbondale
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Variable analysis
independent variables
- Use of anti-Pus10 (HPA049582, Sigma) antibody
- Use of another commercial anti-Pus10 (HPA044736, Sigma) antibody
- Overexpression of Pus10 using anti-HisTag antibody (2365S, Cell Signaling)
- Immunoblotting results shown in Figure 3C and Supplemental Figure S5B
- QPCR results (not explicitly mentioned)
- Immunoblotting and qPCR procedures were done exactly as described previously (Jana et al. 2017)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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