Affinity Purification of Protein Complexes in C. elegans - Caenorhabditis Elegans: Molecular Genetics and Development - page 289

Fig. 1 Experimental outline for protein complex identification in C. elegans. Wild-type or LAP-tagged
strain is grown on NGM plates until larvae are starved. With starved larvae an unsynchronized starter
culture is inoculated. Embryos are isolated by bleaching and hatched in the absence of food to obtain
starved L1 larvae. Starved L1 larvae are used to set up six synchronized liquid cultures. After several
rounds of synchronized liquid culture, when sufficient amounts of worms/embryos are obtained, the
extract is prepared and protein complexes are purified by immunoprecipitation and analyzed by mass
spectrometry. Approximate time for each experiment is indicated. (See color plate.)
#44938) and also blocked by adding an unrelated fusion protein preparation that
harbors similar contaminants. For both immunofluorescence and immunoblotting,
we typically use 0.5-1 m g/mL affinity-purified antibody in the primary antibody
incubation step. To deplete contaminating anti-bacterial protein antibodies, we
incubate 500 m Lof10 m g/mL affinity purified antibody (diluted in AbDil: PBS