Cell extracts were prepared in RIPA buffer (Pierce) supplemented with Protease Inhibitor Cocktail (Sigma) and separated on 4–12% gradient sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gels. Western blotting was performed as described previously (6 (link)). Anti-P54nrb (sc-376865) and anti-PSF (sc-374502) antibodies were purchased from Santa Cruz Biotechnology. Anti-PSPC1 (ab104238), anti-hnRNP K (ab32969), anti-TCP1-β (ab92746) and anti-FUS (ab23439) were purchased from Abcam. Anti-GAPDH (G8795) and anti-γ-Tubulin (T6557) were purchased from Sigma. Anti-phospho-Cyclin D1 (Thr286) (3300) was purchased from Cell Signaling.
Protocol detail
Western Blot Analysis of Nuclear Proteins
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Antibodies
Buffer
Cell extracts
Cyclin d1
Gapdh
Gels
Hnrnp k
Protease inhibitor
Ripa
Sds page
Tubulin
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Variable analysis
independent variables
- Cell extracts were prepared in RIPA buffer (Pierce) supplemented with Protease Inhibitor Cocktail (Sigma)
- Separation of proteins on 4–12% gradient sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gels
- Western blotting
- GAPDH
- γ-Tubulin
- Anti-P54nrb (sc-376865)
- Anti-PSF (sc-374502)
- Anti-PSPC1 (ab104238)
- Anti-hnRNP K (ab32969)
- Anti-TCP1-β (ab92746)
- Anti-FUS (ab23439)
- Anti-phospho-Cyclin D1 (Thr286) (3300)
- Not explicitly mentioned
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