Anti neun antibody mab377

To detect neuronal apoptotic cells, sections were treated with 0.1 M citrate buffer at 90 °C (pH 6.0) for antigen retrieval. TUNEL assay was used with an in-situ cell death detection kit (Roche, Mannheim, Germany) followed by incubating with mouse monoclonal primary anti-NeuN antibody (MAB377; Millipore, US) at 4 °C overnight. Sections were then incubated with donkey anti-mouse IgG Alexa Fluor® 594-conjugated secondary antibody (Thermofisher, US) for 30 mins. Sections were mounted by slow fade® anti-fade DAPI reagent (Lifetech, US).
To quantify the degree of apoptotic cell death in the cortex and hippocampus, ten coronal sections from rostro-to-caudal region of the cortex and hippocampus were examined and TUNEL-positive cells were manually counted in a defined sampling region. One sampling region of interest from the hippocampal CA1 and two sampling regions in the cortex were defined. Only TUNEL-label cells colocalized with condensed nuclei staining were counted. The number of TUNEL-positive cells was the sum of apoptotic cell counts from all sample fields in the quantified sections of one embedded block. The number of TUNEL-positive cells reported was the mean of n number in each group.

Chitosan (85% deacetylation, molecular weight 10⁵ Da) was purchased from Zhejiang Jinke Biochemistry (Zhejiang, China). Rhodamine B isothiocyante (RBITC) was obtained from Sigma-Aldrich (USA). Sodium tripolyphosphate (TPP) was purchased from Shijiazhuang Shinearly Chemicals (China). Polysorbate 80 was obtained from Sinopharm Chemical Reagent (China). The following kits were purchased from Beyotime (Haimen, China): H₂O₂ assay kit (S0038), total glutathione assay kit (S0052), and lipid peroxidation MDA assay kit (S0131). Anti-NeuN antibody (MAB377) was purchased from Millipore (MA, USA). Rabbit anti-GFAP was obtained from Wuhan Boster Company (Wuhan, China).