Indirect Immunofluorescence

The indirect immunofluorecence procedure is performed in the dark to prevent bleaching of the label.

1. Specific immunoreaction: Apply primary mouse anti-P-tubulin antibody (1/50 dilution in 1X PBS) for 30 min (see Note 14 ), followed by three quick (1 min each) washes in 1X PBS.

2. Revelation by the secondary (labeled) antibody: incubate with donkey TRITC-labeled anti-mouse F(ab')2 fragments for 30 min (seeNote 1 4) and wash three times in 1X PBS.

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