Antibody Staining Protocol for c-Fos and Netrin-G1

Primary antibodies used in this study included rabbit polyclonal antibodies for netrin-G112 (link), rabbit polyclonal anti-c-Fos (Ab-5, Calbiochem), rabbit polyclonal anti-GABA (Sigma), mouse monoclonal anti-NeuN (Chemicon), rabbit polyclonal anti-ß-galactosidase (MP Biomedicals), and mouse monoclonal anti-ß-galactosidase (Promega). The secondary antibodies used for immunofluorescence were Alexa Fluor 488 and 546 donkey anti-mouse or rabbit IgG (Molecular Probes). Biotinylated goat anti-rabbit IgG and a Vectastain ABC kit (Vector Laboratories) were used for c-Fos staining. Mice for c-Fos analysis were firstly exposed to EPM or FC training and then sacrificed and perfused 100 minutes later. The c-Fos staining was performed as described previously59 (link). Rabbit polyclonal antibodies against Netrin-G1 were used as described previously12 (link).

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Zhang Q., Sano C., Masuda A., Ando R., Tanaka M, & Itohara S. (2016). Netrin-G1 regulates fear-like and anxiety-like behaviors in dissociable neural circuits. Scientific Reports, 6, 28750.

Publication 2016

anti-GABA mouse monoclonal anti-NeuN Alexa Fluor 488 546 donkey anti-mouse or rabbit IgG Vectastain ABC kit

Alexa fluor 488 Anti igg Antibodies C fos Donkey Gaba Galactosidase Goat Immunofluorescence Mice Molecular probes Netrin Netrin g1 Promega Rabbit Vector

Corresponding Organization :

Other organizations : RIKEN Center for Brain Science

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Variable analysis

independent variables

Exposure to EPM (elevated plus maze)
FC (fear conditioning) training

dependent variables

C-Fos staining

control variables

Time between exposure/training and sacrifice (100 minutes)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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