Alexa 647 conjugated goat anti guinea pig igg antibody

The fifth series of the sections through the Vc was incubated overnight with a mixture of 1 μg/ml guinea pig anti-FG antibody (Protos Biotech Corporation), 0.5 μg/ml rabbit anti-TMR antibody(Invitrogen), and 1 μg/ml goat anti-CGRP antibody (ab36001; Abcam) in PBS-XCD. After three washes with PBS, the sections were treated with 10 μg/ml biotinylated donkey anti-goat IgG (AP180B, Millipore, Temecula, CA, USA). The sections were then further incubated overnight with a mixture of 10 μg/ml Alexa 647-conjugated goat anti-guinea pig IgG antibody (Invitrogen), 10 μg/ml Alexa 594-conjugated donkey anti-rabbit IgG antibody (Invitrogen), and 10 μg/ml Fluorescein Avidin D (Vector) in PBS that contained 5% (v/v) normal donkey serum.
The sixth sections, which contained the Vsp of the rats injected with FG and TMR, were used to conduct the control experiments for immunofluorescence histochemistry, in which CGRP antibody was omitted. Under these conditions, no immunoreactivity for the omitted antibody was observed.

The last sections through the Vc of the rats in the first group and the sections through the Vc from the second group injected with FG, TMR, and formalin were also processed for triple-immunofluorescence histochemistry for Fos, TMR and FG. Briefly, the sections were incubated with (1) a mixture of 1 μg/ml mouse anti-Fos antibody (ab 208,942, Abcam, Cambridge, MA, USA), 1 μg/ml guinea pig anti-FG antibody (NM-101, Protos Biotech Corporation) and 0.5 μg/ml rabbit anti-TMR antibody (A-6397, Invitrogen) in PBS that contained 0.3% (v/v) Triton X-100, 0.25% (w/v) λ-carrageenan, and 3% (v/v) donkey serum (PBS-XCD) overnight at room temperature; (2) 10 μg/ml biotinylated donkey anti-mouse IgG (AP192B, Millipore, Temecula, CA, USA); and (3) a mixture of 10 μg/ml Alexa 647-conjugated goat anti-guinea pig IgG antibody (A-21450, Invitrogen), 10 μg/ml Alexa594-conjugated donkey anti-rabbit IgG antibody (A-21207, Invitrogen), and 10 μg/ml Fluorescein Avidin D (A-2001; Vector) in PBS that contained 3% (v/v) normal donkey serum.
Another series of sections from the second group was used to conduct the control experiments for immunofluorescence histochemistry, in which the Fos antibody was omitted. Under these conditions, no immunoreactivity for the omitted antibody was observed.