Immunostaining of Drosophila CNS

The central nervous system (CNS) was dissected from flies in PBS, and fixed in 4% paraformaldehyde dissolved in PBS for 60 min. Immunostaining was carried out as described previously [16 (link)], using the following antibodies at the indicated dilutions: rabbit anti-GFP (Sigma, A6455; 1:500), mouse Anti-Brp (nc82) (DSHB, 1:10), guinea pig anti-FruMale [48 (link)] (1:500) and rabbit anti-Camta (this study, 1:200). The secondary antibodies used were as follows: Alexa488-conjugated goat anti-rabbit IgG (Thermo Fisher, A-11034) at 1:500, Alexa546-conjugated goat anti-mouse, rabbit or guinea pig IgG (Thermo Fisher, A-11003, A-11035 or A-11074) at 1:500, and Alexa647-conjugated goat anti-mouse IgG (Thermo Fisher, A32723) at 1:500. Stacks of optical sections were obtained with a Zeiss LSM 510 META confocal microscope and were processed with Image J software.

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Sato K., Ahsan M.T., Ote M., Koganezawa M, & Yamamoto D. (2019). Calmodulin-binding transcription factor shapes the male courtship song in Drosophila. PLoS Genetics, 15(7), e1008309.

Publication 2019

rabbit anti-GFP Alexa488-conjugated goat anti-rabbit IgG Alexa647-conjugated goat anti-mouse IgG LSM 510 META confocal microscope

Alexa647 Anti igg Antibodies Central nervous system Confocal microscope Dilutions Flies Goat Guinea pig Mouse Paraformaldehyde Rabbit Sections

Corresponding Organization : Tohoku University

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