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Walton D. Jones added ihc.tex
almost 9 years ago
Commit id: 38fc7df51943e23827c41a1bc19a67b07f87bf42
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\subsection*{Immunohistochemistry}
Adult brain of female flies was dissected in PBS and fixed with 4\% PFA for 30 minutes at room temperature.
Fixed brain samples were washed with PAT3 (0.5\% TritonX-100, 0.5\% BSA in PBS) for 15 min 3 times and incubated in 5\% normal goat serum for 1-2 hours at room temperature.
After blocking, samples were incubated with primary antibody overnight at $4\degree$C.
Samples were washed with PAT3 for 1 hour twice at room temperature and incubated with secondary antibody overnight at $4\degree$C.
After washed with PAT3 for 1 hour twice, brain samples were mounted with vectashield solution (H-1000, Vector Laboratories, Inc.).
Primary antibody used: rabbit anti-GFP antibody 1: 1000(A11122, Invitrogen); monoclonal anti-bruchpilot antibody (nc82, DSHB).
Secondary antibody used: Goat anti-rabbit Alexa488 1:400 (A11008, Invitrogen); Goat anti-mouse Alexa568 1:400 (A11031, Invitrogen).
