Non-Abcam antibody was used: goat anti rabbit AlexaFluor555 Host species: Goat Clonality: Polyclonal Conjugation: Alexa Fluor® 555
Cells were plated on an uncoated cover slip. One day after plating cells were washed twice with PBS and fixed with 4% PFA incl. 4% sucrose for 15 minutes at room temp. Cell were permeabilized with 0.5% saponin in PBS for 45min at 4°C. Blocking was carried out with 10% fetal calf serum in PBS-saponin during first and second antibody incubation. After additional washing cover splips were mounted on glass slides with ProLong® Gold antifade reagent with DAPI (Invitrogen). Antibody can probably be diluted further than 1:200.
Abcam user community
Submitted Sep 03 2010
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