Table 1.
Antibody manufacturer and dilutions used for immunohistochemistry
| Primary Antibody, Dilution | Secondary Antibody, Dilution | Dye, Dilution |
|---|---|---|
| All Neurons | ||
| Ms monoclonal HuC/HuD, 1:200 Molecular Probes, Eugene, OR A21271 |
Dk anti Ms IgG Dylight 488, 1:300 Jackson mmunoResearch, West Grove, PA 715-485-150 |
N/A |
| Inhibitory Motor Neurons | ||
| Rb polyclonal VIP, 1:1000 Abcam, Cambridge, MA Ab43841 |
Biotinylated Gt anti Rb IgG, 1:300 Vector Labs, Burlingame, CA BA1000 |
Streptavidin Alexa Fluor 488 conjugate, 1:600; Jackson ImmunoResearch, West Grove, PA 016-540-084 |
| Rb polyclonal nNOS, 1:500 Invitrogen, Camarillo, CA #61-7000 |
Biotinylated Gt anti Rb IgG, 1:300 Vector Labs, Burlingame, CA BA1000 |
Streptavidin Cy3 conjugate, 1:600 Life Technologies, Grand Island, NY SA1010 |
| Sheep polyclonal nNOS, 1:500 Abcam, Cambridge, MA Ab6175 |
Dk anti Sheep IgG Dylight 594, 1:300 Jackson ImmunoResearch, West Grove, PA 715-485-150 |
N/A |
| Excitatory Motor Neurons | ||
| Gt polyclonal ChAT, 1:200 Millipore, Billerica, MA AB144P |
Biotinylated Rb anti Gt IgG, 1:300 Vector Labs, Burlingame, CA BA5000 |
N/A |
| Rt monoclonal Substance P, 1:150 Millipore, Billerica, MA MAB356 |
Biotinylated Gt anti Rt, 1:300 Invitrogen, Camarillo, CA #62–9540 |
Streptavidin Cy3 conjugate, 1:600 Life Technologies, Grand Island, NY SA1010 |
| Sensory Neurons | ||
| Rabbit polyclonal anti-Calbindin D-28k, 1:1250 CH-1723, Marly 1 (Switzerland) CB 38 |
Biotinylated Gt anti Rb IgG, 1:300 Vector Labs, Burlingame, CA BA1000 |
Streptavidin Cy3 conjugate, 1:600 Life Technologies, Grand Island, NY SA1010 |
| Rb polyclonal CGRP, 1:200 Sigma Aldrich, St. Louis, MO # c8198 |
Biotinylated Gt anti Rb IgG, 1:300 Vector Labs, Burlingame, CA BA1000 |
Streptavidin Cy3 conjugate, 1:600 Life Technologies, Grand Island, NY SA1010 |
| Apoptosis | ||
| Rb polyclonal anti-cleaved Caspase-3, Cell Signaling Technology (ASP 175) |
Dk anti rabbit IgG Dylight 488, 1:300 | N/A |
Ms: Mouse, Dk: Donkey, Rb: Rabbit, Gt: Goat, Rt: Rat