Table 1.
Antisera used in each group.
| Methods | Purpose | Primary antiserum | Secondary antiserum | Tertiary antiserum |
|---|---|---|---|---|
| Light microscopy | NTR2/BDA/FG | Rabbit anti-NTR2 (1:1000, Millipore, Temecula, CA, USA) Guinea pig anti-FG (1:100, Protos Biotech, New York, NY, USA) | Alexa594 donkey anti-rabbit (1:500, Invitrogen, Camarillo, CA, USA) Alexa488 goat anti- guinea pig (1:500, Invitrogen) Alexa647 avidin (1:1000, Invitrogen) | |
| FG/BDA/NT | Guinea pig anti-FG (1:100, Protos Biotech) Rat anti-NT (1:200, Protos Biotech) | Alexa488 goat anti-guinea pig (1:500, Invitrogen) Alexa594 goat anti-rat (1:500, Invitrogen) Alexa647 avidin (1:1000, Invitrogen) | ||
| NTR2/BDA/NT | Rabbit anti-NTR2 (1:1000, Millipore) Rat anti-NT (1:200, Protos Biotech) | Alexa488 donkey anti-rabbit (1:500, Invitrogen) Alexa594 goat anti- rat (1:500, Invitrogen) Alexa647 avidin (1:1000, Invitrogen) | ||
| NTR2/NT/FG | Rabbit anti-NTR2 (1:1000, Millipore) Rat anti-NT (1:200, Protos Biotech) Guinea pig anti-FG (1:100, Protos Biotech) | Biotinylated donkey anti-rabbit (1:500, Millipore) Alexa594 goat anti-rat (1:500, Invitrogen) Alexa488 goat anti-guinea pig (1:500, Invitrogen) | Alexa647 avidin (1:1000, Invitrogen) | |
| NTR2/NT/5-HT | Rabbit anti-NTR2 (1:1000, Millipore) Rat anti-NT (1:200, Protos Biotech) Goat anti-5-HT (1:500, Immunostar, Hudson, WI, USA) | Biotinylated donkey anti-rabbit (1:500, Millipore) Cy3 donkey anti-rat (1:500, Millipore) Alexa647 donkey anti-goat (1:500, Invitrogen) | FITC-avidin (1:1000, Vector Labs) | |
| Electron microscopy | NT/NTR2/HRP* | Rabbit anti-NTR2 (1:1000, Millipore) Rat anti-NT (1:200, Protos Biotech) | Biotinylated donkey anti-Rat IgG (1:200, Millipore) Goat anti-rabbit IgG conjugated to 1.4-nm gold particles (1:100; Nanoprobes) | Avidin-biotinylated peroxidase complex (1:50, Vector Labs) |
| NTR2/BDA/HRP* | Rabbit anti-NTR2 (1:1000, Millipore) | Goat anti-rabbit IgG conjugated to 1.4-nm gold particles (1:100; Nanoprobes) | Avidin-biotinylated peroxidase complex (1:50, Vector Labs) | |
| NT/BDA/HRP* | Rat anti-NT (1:200, Protos Biotech) | Goat anti-rat IgG conjugated to 1.4-nm gold particles (1:100; Nanoprobes) | Avidin-biotinylated peroxidase complex (1:50, Vector Labs) |
*The DAB reaction utilized for the demonstration of HRP in each group was performed before the immuno-electron microscopy procedures.