Sections from the hypothalamus were processed for Fos immunohistochemistry as done previously (Mahler and Aston-Jones, 2012 (link)). Briefly, every 6th section through the hypothalamus was selected to visualize orexin neurons throughout the longitudinal axis of the lateral hypothalamus. Sections were incubated in a rabbit anti-Fos primary antibody overnight (1:1000; Millipore ABE457), followed by 2 hours in a donkey anti-rabbit secondary (1:500, Jackson ImmunoResearch Laboratories, West Grove, PA). The same sections from the hypothalamus were then processed for Orx-A immunohistochemistry in a goat anti-Orx A primary antibody overnight (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA) and then incubated for 2 hours in a donkey anti-goat secondary antibody (1:500, Jackson ImmunoResearch Laboratories, West Grove, PA).
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Fos and Orexin Immunohistochemistry in Hypothalamus
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Anti antibody
Axis
Donkey
Goat
Hypothalamus
Immunohistochemistry
Lateral hypothalamus
Neurons
Orexin
Rabbit
Corresponding Organization : Rutgers, The State University of New Jersey
Other organizations : Furman University, Stanford University, Florey Institute of Neuroscience and Mental Health, University of Melbourne
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Variable analysis
independent variables
- Sections from the hypothalamus
- Fos immunohistochemistry
- Orx-A immunohistochemistry
- Every 6th section through the hypothalamus was selected to visualize orexin neurons throughout the longitudinal axis of the lateral hypothalamus
- Sections were incubated in a rabbit anti-Fos primary antibody overnight (1:1000; Millipore ABE457)
- Sections were then incubated for 2 hours in a donkey anti-rabbit secondary (1:500, Jackson ImmunoResearch Laboratories, West Grove, PA)
- Sections were then processed for Orx-A immunohistochemistry in a goat anti-Orx A primary antibody overnight (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA)
- Sections were then incubated for 2 hours in a donkey anti-goat secondary antibody (1:500, Jackson ImmunoResearch Laboratories, West Grove, PA)
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