We conducted immunohistochemical analyses on the coronal brain slices to assess the expressions of glial fibrillary acidic protein (GFAP), ionized calcium-binding adapter molecule 1 protein (Iba-1), and neuron-specific nuclear protein (NeuN) after blast exposure. The expressions of GFAP, Iba-1, and NeuN are associated with the distributions of astrocytes, microglia, and neuronal cells, respectively21 (link).
At FD Neurotechnologies, the brain slices were incubated with primary antibodies to detect GFAP (#556330; BD Biosciences, San Jose, CA; 1:350 dilution), Iba-1 (#019-19741; Wako Chemicals, Richmond, VA; 1:350 dilution), and NeuN (#ABN90P; Millipore Sigma, Burlington, MA; 1:250 dilution). This was followed by incubation in PBS containing the following secondary reporter antibodies with a fluorescent tag: Alexa Fluor 488 donkey anti-mouse (#A21202; Thermo Fisher Scientific, Waltham, MA; 1:250 dilution), Alexa Fluor 594 donkey anti-rabbit (#A21207; Thermo Fisher Scientific; 1:250 dilution), and Alexa Fluor 405 streptavidin conjugate (#S32351; Thermo Fisher Scientific; 1:200 dilution) for the detection of GFAP, Iba-1, and NeuN primary antibodies, respectively. The stained coronal slices were shipped back to their respective laboratories of origin (WRAIR or NJIT) for quantification analyses.
At FD Neurotechnologies, the brain slices were incubated with primary antibodies to detect GFAP (#556330; BD Biosciences, San Jose, CA; 1:350 dilution), Iba-1 (#019-19741; Wako Chemicals, Richmond, VA; 1:350 dilution), and NeuN (#ABN90P; Millipore Sigma, Burlington, MA; 1:250 dilution). This was followed by incubation in PBS containing the following secondary reporter antibodies with a fluorescent tag: Alexa Fluor 488 donkey anti-mouse (#A21202; Thermo Fisher Scientific, Waltham, MA; 1:250 dilution), Alexa Fluor 594 donkey anti-rabbit (#A21207; Thermo Fisher Scientific; 1:250 dilution), and Alexa Fluor 405 streptavidin conjugate (#S32351; Thermo Fisher Scientific; 1:200 dilution) for the detection of GFAP, Iba-1, and NeuN primary antibodies, respectively. The stained coronal slices were shipped back to their respective laboratories of origin (WRAIR or NJIT) for quantification analyses.
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