Immunohistochemistry of c-Fos in Mouse Brain

Mice were deeply anesthetized with Euthasol and transcardially perfused with 4% paraformaldehyde in phosphate-buffered saline (PBS), pH 7.4. Coronal brain slices (35 μm) were prepared, and a standard immunohistochemistry protocol was followed (17 (link)). The primary antibody used was rabbit anti–c-Fos (1:1000; Abcam, ab-222699 RRID: AB_2891049). Alexa Fluor secondary antibodies (633-goat anti-rabbit; 1:1000; Life Technologies, A21052, RRID: AB_2535719) were used to visualize the signal, and images were acquired by confocal microscopy (Zeiss, LSM880) (17 (link)).

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Huang Z., Liu L., Zhang J., Conde K., Phansalkar J., Li Z., Yao L., Xu Z., Wang W., Zhou J., Bi G., Wu F., Seeley R.J., Scott M.M., Zhan C., Pang Z.P, & Liu J. (2022). Glucose-sensing glucagon-like peptide-1 receptor neurons in the dorsomedial hypothalamus regulate glucose metabolism. Science Advances, 8(23), eabn5345.

Publication 2022

ab-222699 A21052 LSM880

Antibodies Antibody Brain Confocal microscopy Goat Immunohistochemistry Mice Paraformaldehyde Phosphate Rabbit Saline

Corresponding Organization : University of Science and Technology of China

Other organizations : Rutgers, The State University of New Jersey, Johnson University, University of Michigan–Ann Arbor, University of Virginia, Tsinghua University, National Institute of Biological Sciences, Beijing

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Variable analysis

independent variables

Euthasol (anesthetic)

dependent variables

Expression of c-Fos protein (as measured by immunohistochemistry)

control variables

Paraformaldehyde concentration (4% in phosphate-buffered saline)
PH of phosphate-buffered saline (7.4)
Brain slice thickness (35 μm)
Antibody concentrations (primary antibody: 1:1000; secondary antibody: 1:1000)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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