For Western blotting experiments, we collected individual worms by hand and boiled them in Laemmli protein sample loading buffer prior to gel separation. Specific proteins were analyzed by western blotting with ECL detection (GE Healthcare) of HRP-coupled secondary antibodies (Jackson ImmunoResearch). Worm protein extracts for sucrose gradients and ribosome immunoprecipitations were made as described (5 (link)). The sucrose gradient centrifugation and fractionation was conducted as previously described (10 (link)).
Protocol detail
Protein Extraction and Western Blotting for Worms
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Antibodies
Centrifugation
Fractionation
Immunoprecipitations
Laemmli buffer
Proteins
Ribosome
Sucrose
Worm protein
Worms
Corresponding Organization : Martin Luther University Halle-Wittenberg
Other organizations : Max Planck Institute of Biochemistry
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Variable analysis
independent variables
- Collecting individual worms by hand
- Specific proteins analyzed by western blotting with ECL detection
- Worm protein extracts for sucrose gradients and ribosome immunoprecipitations
- Boiling worms in Laemmli protein sample loading buffer prior to gel separation
- Sucrose gradient centrifugation and fractionation as previously described
- Positive controls not explicitly mentioned
- Negative controls not explicitly mentioned
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