Alexa 647 goat antibody to rabbit igg

Single and double immunohistochemical reactions were performed as described previously (Garcia-Moreno et al., 2012 (link)) using the following primary antibodies: Rabbit antibody to EGFP (Molecular Probes, A11122, 1:1000), mouse antibody to EGFP (Abcam, ab1218, 1:1000), chick antibody to EGFP (Aves GFP 1020, 1:10000), rabbit anti Nurr1 (Santa Cruz, sc-991, 1:400), rabbit antibody to Dbx1 (1:200; kind gift by Prof. Nakagawa, Univ. Minessota, United States), rabbit antibody to Tbr1 (Chemicon, AB9616, 1:1,000), rat antibody to Ctip2 (Abcam, ab18465, 1:500), rabbit antibody to dsRed2 (Takara, 632475, 1:1000), and rabbit antibody to Pax6 (Covance, PRB-278P, 1:200). Dbx1 immunostaining required antigen retrieval with citrate acid.
For secondary antibodies (all 1:1000), we used Alexa 568 goat antibody to rabbit IgG (Molecular Probes, A11011), Alexa 647 goat antibody to rabbit IgG (Molecular Probes, A21245), Alexa 488 goat antibody to rabbit IgG (Molecular Probes, A11034), Alexa 488 goat antibody to mouse IgG (Molecular Probes, A11001), Alexa 568 antibody to mouse IgG (Molecular Probes, A11004), Alexa 568 goat antibody to rat IgG (Molecular Probes, A11077), and Alexa 488 goat antibody to chicken (Invitrogen, A11039).

Embryonic chick brains were fixed by immersion in PFA (4% paraformaldehyde, PFA, diluted in phosphate-buffered saline 0.1 M– PBS, pH 7.3). Brains were transferred to PBS 6 h after fixation. Brains were sectioned in the coronal plane at 50–70-μm thickness in a vibrating microtome (Leica VT1000S). Single and double immunohistochemical reactions were performed as described previously⁵² using the following primary antibodies: rabbit antibody to histone H3-phospho S10 (Abcam ab47297; 1:1000), mouse antibody to LHX5 (DSHB;1:30) and mouse antibody to Meis2 (DSHB; 1:30). For secondary antibodies (all 1:1000), we used Alexa 647 goat antibody to rabbit IgG (Molecular Probes, A32733) and Alexa 488 goat antibody to mouse IgG (Molecular Probes, A11001). Sections were counterstained with DAPI.