|
Fos/TH Immunohistochemistry |
|
Fos |
Serum |
Normal goat serum. Pel-Freez Biologicals, Rogers, AR |
Primary |
c-Fos (4): rabbit, sc-52. 1:10,000, 0.02μg IgG/ml solution, Santa Cruz Biotech, Santa Cruz, CA |
Secondary |
Biotinylated goat anti-rabbit IgG (H+L). 1:500, 3μg IgG/ml solution, Vector Laboratories, Burlingame, CA |
TH |
Serum |
Normal goat serum. Pel-Freez Biologicals, Rogers, AR |
Primary |
Mouse anti-TH monoclonal antibody. 1:2,000, Millipore-Chemicon, Billerica, MA |
Secondary |
Biotinylated goat anti-mouse IgG (H+L). 1:500, 3μg IgG/ml solution, Vector Laboratories, Burlingame, CA |
Fos/orexin Immunohistochemistry |
|
Fos |
Serum |
Normal donkey serum. Jackson ImmunoResearch Laboratories, West Grove, PA |
Primary |
c-Fos (4): rabbit, sc-52. 1:10,000, 0.02μg IgG/ml solution, Santa Cruz Biotech, Santa Cruz, CA |
Secondary |
Biotinylated donkey anti-rabbit IgG (H+L). 1:500, 2.4μg IgG/ml solution, Jackson ImmunoResearch, West Grove, PA |
Orexin |
Serum |
Normal horse serum, S-2000. Vector Laboratories, Burlingame, CA |
Primary |
Orexin-A (C-19): goat, sc-8070, 1:5,000, 0.04μg IgG/ml solution, Santa Cruz Biotech, Santa Cruz, CA |
Secondary |
Biotinylated horse anti-goat IgG (H+L), 1:500, 3μg IgG/ml solution, Vector Laboratories, Burlingame, CA |
Other reagents |
|
Avid :Biotin Complex |
Peroxidase- Vectastain ABC Kit PK-6100, Vector Laboratories, Burlingame, CA |
Enzyme Substrate |
Peroxidase- Vector SG Substrate Kit SK-4700, Vector Laboratories, Burlingame, CA |
Enzyme Substrate |
Peroxidase- 3,3′ Diaminobenzidine tetrahydrochloride, Sigma-Aldrich, St. Louis, MO |