Table 2.
Primary and secondary antibodies and avidins used for localizing various antigens by immunofluorescence
| Primary antibody | Host | Antibody type | Dilution factor | Manufacturer | Catalog no | Lot no |
|---|---|---|---|---|---|---|
| ChAT | Rabbit | Monoclonal IgG | 1:1000—5000 | Abcam, Cambridge, UK | ab178850 | EPR16590 |
| GAD67 | Mouse | Monoclonal IgG | 1:1000 | Millipore | MAB5406 | |
| GFP | Rabbit | Polyclonal | 1:2000 | Molecular Probes, Eugene, USA | A6455 | 1,650,113 |
| GFP | Chicken | Polyclonal | 1:400 | AvES labs inc., Tigard, USA | GFP-1020 | GFP697986 |
| PV27 | Rabbit | Polyclonal IgG | 1:3000 | Swant, Inc., Marly, Switzerland | ||
| PV235 | Mouse | Monoclonal IgG | 1:1000—5000 | Swant, Inc., Marly, Switzerland | 10-11F | |
| Serotonin (5-HT) | Rabbit | Polyclonal | 1:1000 | Immuno nuclear corporation, Stillwater, USA | 43H2TR | 8,344,016 |
| Tyrosine hydroxylase (TH) | Mouse | Monoclonal IgG | 1:1000—3,000 | Immunostar, Hudson, USA | 22,941 | 1,602,001 |
| VGLut1 | Rabbit | Polyclonal | 1:1000—3000 | Synaptic systems | 135,403 | AB_887883 |
| VGlut2 | Rabbit | Polyclonal | 1:1000—30000 | Synaptic systems | 135,403 | AB_2315570 |
| Biotinylated secondary antibody | ||||||
| Biotinylated anti-mouse | Horse | IgG (H + L) | 1:250 | Vector laboratories, Burlingame, Ca, USA | BA-2000 | ZA0409/Z0715 |
| Biotinylated anti-rabbit | Goat | IgG (H + L) | 1:250 | Vector laboratories, Burlingame, Ca, USA | BA-1000 | ZB1007 |
| Fluorochrome-conjugated secondary antibody | ||||||
| Alexa fluor 488 anti-mouse | Donkey | IgG (H + L) | 1:200 | Jackson immuno research, Suffolk, UK | A21202 | 1,226,927 |
| Alexa fluor™ 488 anti-rabbit | Donkey | IgG (H + L) | 1:200 | Jackson immuno research, Suffolk, UK | A21206 | 1,834,802 |
| Alexa fluor 488 anti-chicken | Donkey | IgG (H + L) | 1:200 | Jackson immuno research, Suffolk, UK | 703-225-155 | |
| Cy3 anti-chicken | Donkey | IgG (H + L) | 1:200 | Jackson immuno research, Suffolk, UK | 703-165-155 | 120,431 |
| Cy3 anti-mouse | Donkey | IgG (H + L) | 1:200 | Jackson immuno research, Suffolk, UK | ||
| Cy5 anti-rabbit | Donkey | IgG (H + L) | 1:200 | Jackson immuno research, Suffolk, UK | ||
| Antibody detection | ||||||
| Streptavidine Cy2 | 1:200 | Jackson immuno research, Suffolk, UK | 016-540-084 | |||
| Streptavidine Cy3 | 1:200 | Jackson immuno research, Suffolk, UK | 016-160-084 | |||