rabbit polyclonal anti ncx  (Alomone Labs)

Journal: Journal of Molecular Histology

Article Title: Neuroprotection in neonatal Hypoxia-ischaemia: melatonin targets NCX1 to inhibit mitochondrial autophagy via the PINK1-Parkin pathway

doi: 10.1007/s10735-025-10601-5

Figure Lengend Snippet: Immunohistochemical and simple Western blot analysis to evaluate the effect of melatonin on NCX1 expression activity. a Immunohistochemistry was used to assess the expression levels of NCX1 in neonatal rats at 7, 14, and 21 days. In the Sham group, normal NCX1 protein expression was observed in the brain. Compared with the Sham group, the HI group showed significantly greater NCX1-positive staining at 7 days of age after the brain injury. However, the positive results for NCX1 were diminished at 14 and 21 days of age. In contrast, strong positive expression was demonstrated in the Mel group at 14 and 21 days of age. Compared with the Mel group, the SN6 group exhibited high expression at 7 days of age and low expression at 14 and 21 days of age; scale bar = 20 µm, b Average optical density (AOD) of NCX1 staining in each group (n = 6), c – d NCX1 protein expression exhibited dynamic changes in the HI group, with an increase observed on postnatal Day 7 compared with that in the Sham group but a significant decrease on Days 14 and 21. This trend was reversed in the Mel group, whose expression was lower than that in the HI group on Day 7 but significantly greater on Days 14 and 21. In the SN6 group, expression was upregulated on Day 7 but decreased at subsequent time points (n = 4), e There were no significant differences in the expression of NCX1 or autophagy-related proteins among the S, S + M, and S + M + N groups (n = 3); the data represent the mean ± SEM; ns p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The primary antibodies used included: Rabbit anti-NCX1 polyclonal antibody (1:20, 0.2 μg/μL, #28447-1-AP; Proteintech, Wuhan, China) Rabbit anti-PLP monoclonal antibody (1:10, 2 μg/μL, EPR23504-106, #ab254363; Abcam, Cambridge, UK) Rabbit anti-LC3B monoclonal antibody (1:100, 0.2 μg/μL, EPR18709 , #ab192890; Abcam) Mouse anti-MAG monoclonal antibody (1:20, 2 μg/μL, sc-166849; Santa Cruz Biotechnology, Dallas, TX, USA) Mouse anti-PINK1 monoclonal antibody (1:100, 2 μg/μL, sc-517353; Santa Cruz Biotechnology) Mouse anti-Parkin monoclonal antibody (1:20, 2 μg/μL, sc-32282; Santa Cruz Biotechnology)

Techniques: Immunohistochemical staining, Simple Western, Expressing, Activity Assay, Immunohistochemistry, Staining

Journal: Journal of Molecular Histology

Article Title: Neuroprotection in neonatal Hypoxia-ischaemia: melatonin targets NCX1 to inhibit mitochondrial autophagy via the PINK1-Parkin pathway

doi: 10.1007/s10735-025-10601-5

Figure Lengend Snippet: Immunohistochemical and simple Western blot analysis to evaluate the effect of melatonin on NCX1 expression activity. a Immunohistochemistry was used to assess the expression levels of NCX1 in neonatal rats at 7, 14, and 21 days. In the Sham group, normal NCX1 protein expression was observed in the brain. Compared with the Sham group, the HI group showed significantly greater NCX1-positive staining at 7 days of age after the brain injury. However, the positive results for NCX1 were diminished at 14 and 21 days of age. In contrast, strong positive expression was demonstrated in the Mel group at 14 and 21 days of age. Compared with the Mel group, the SN6 group exhibited high expression at 7 days of age and low expression at 14 and 21 days of age; scale bar = 20 µm, b Average optical density (AOD) of NCX1 staining in each group (n = 6), c – d NCX1 protein expression exhibited dynamic changes in the HI group, with an increase observed on postnatal Day 7 compared with that in the Sham group but a significant decrease on Days 14 and 21. This trend was reversed in the Mel group, whose expression was lower than that in the HI group on Day 7 but significantly greater on Days 14 and 21. In the SN6 group, expression was upregulated on Day 7 but decreased at subsequent time points (n = 4), e There were no significant differences in the expression of NCX1 or autophagy-related proteins among the S, S + M, and S + M + N groups (n = 3); the data represent the mean ± SEM; ns p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: The sections were incubated overnight at 4 °C with a rabbit anti-NCX1 monoclonal antibody (Proteintech, #26,835–1-AP, 1:300).

Techniques: Immunohistochemical staining, Simple Western, Expressing, Activity Assay, Immunohistochemistry, Staining