rabbit polyclonal anti sod2  (Proteintech)

Journal: Animal Cells and Systems

Article Title: Epinephrine as a potential driver of oral lichen planus pathogenesis

doi: 10.1080/19768354.2025.2588914

Figure Lengend Snippet: High level of epinephrine induces intracellular oxidative stress in oral keratinocytes. (A) HOK-16B cells treated with PBS or epinephrine for 24 h were subjected to CellROX staining. Nuclear DAPI signal is shown in blue (left). The relative CellROX signal intensities are shown (right). Each symbol means an individual cell. (B) Immunoblot analysis of SOD2 and SESN2 in HOK-16B cells treated with epinephrine for 24 h. β ACTIN was used as loading control. (C) Relative band intensities are shown. (D) Relative mRNA expression of TNFα, IL6, and IL8 in HOK-16B cells treated with epinephrine for 3 h. ** P < 0.01; *** P < 0.001. n.s , not significant.

Article Snippet: After blocking with 5% non-fat dried milk in Tris-buffered saline with TweenTM 20 (TBST) (10 mM Tris, pH 8.0, 150 mM NaCl and 0.5% Tween 20) for 30 min, membranes were incubated with antibodies against phospho STAT3 (1:1000, Cell signaling, Danvers, MA, USA, cat no. 9145), total STAT3 (1:1000, Cell signaling, cat no. 8768), β ACTIN (1:5000, Sigma Aldrich, cat no. A5316), phospho ERK (1:1000, Thermo Fisher Scientific, Waltham, MA, USA, cat no. MA5-15174), total ERK (1:1000, Cell signaling, cat no. 4695), phospho AKT (1:1000, Cell signaling, cat no. 4058), total AKT (1:1000, Cell signaling, cat no. 9272), BCL2 (1:1000, Bioworld Technology, China, cat no. BS1511), SOD2 (1:1000, Cusabio, Houston, TX, USA, cat no. CSB-PA022398LA01HU), and SESN2 (1:1000, Abcam, Cambridge, UK, cat no. ab178518) overnight at 4°C.

Techniques: Staining, Western Blot, Control, Expressing

Journal: bioRxiv

Article Title: Compartmental Differences in the Retinal Ganglion Cell Mitochondrial Proteome

doi: 10.1101/2024.05.07.593032

Figure Lengend Snippet: Proteomic profiling of mitochondria isolated from RGC somatodendritic and axonal compartments. A. Venn diagrams depicting proteins identified in 4 biological replicates for each compartment. Proteins identified in at least 3 of the 4 replicates are bolded and underlined. B. Assigned subcellular localizations of proteins identified in each compartment C. Volcano plot showing the relative abundance of proteins identified in both RGC somatodendritic (Ret) and axonal (ON) compartments. Proteins to the left of the vertical line are more abundant in the somatodendritic compartment and those to the right are more abundant in the axonal compartment. Colored dots represent proteins that underwent immunohistochemical validation: SOD2 (purple), SFXN3 (red), and HADHA (blue). D. Comparison of GFP-normalized protein abundance for SOD2, SFXN3, and HADHA between the somatodendritic and axonal compartments. A two tailed t-test was performed for each comparison. The GFP-normalized protein abundance for each replicate is depicted as an individual data point.

Article Snippet: The following antibodies with specified dilutions were used for immunofluorescence (IF) and western blot (WB) analyses: goat polyclonal anti-GFP ([1:1000 WB, 1:750 IF]; abcam, ab5450), rabbit polyclonal anti-HADHA (1:50 IF; abcam, ab54477), rabbit polyclonal anti-SOD2/MnSOD (1:50 IF; abcam, ab13534), mouse monoclonal anti-NDUFS4 (1:1000 WB; Santa Cruz Biotechnology, sc-100567), mouse monoclonal anti-Tuj1 (1:500 IF; Fisher Scientific, MAB11905), mouse polyclonal anti-VDAC (1:1000 WB; EMD Millipore, AB10527), rabbit polyclonal anti-SFXN3 (1:50 IF; Sigma, HPA008028).

Techniques: Isolation, Immunohistochemical staining, Comparison, Two Tailed Test

Journal: bioRxiv

Article Title: Compartmental Differences in the Retinal Ganglion Cell Mitochondrial Proteome

doi: 10.1101/2024.05.07.593032

Figure Lengend Snippet: Histological validation of compartment-specific protein abundance in RGC mitochondria. A,B. Retinal (A) and optic nerve cross sections (B) from RGC MitoTag mice showing Vglut2-cre -dependent expression of GFP-OMM in RGCs (left panels, green), endogenous expression of proteins of interest (POI; middle panels, magenta), and POI expression solely within GFP-positive mitochondria (right panels, magenta). Protein identities are indicated to the left. Scale bars, 50 µm (A) and 20 µm (B). C. Comparison of GFP-normalized fluorescence intensities of SOD2, SFXN3 and HADHA in retinal and optic nerve sections from RGC MitoTag mice (N=4 replicates for each tissue). A two tailed t-test was performed for each comparison. The GFP-normalized fluorescence intensities for each replicate are depicted as individual data points.

Article Snippet: The following antibodies with specified dilutions were used for immunofluorescence (IF) and western blot (WB) analyses: goat polyclonal anti-GFP ([1:1000 WB, 1:750 IF]; abcam, ab5450), rabbit polyclonal anti-HADHA (1:50 IF; abcam, ab54477), rabbit polyclonal anti-SOD2/MnSOD (1:50 IF; abcam, ab13534), mouse monoclonal anti-NDUFS4 (1:1000 WB; Santa Cruz Biotechnology, sc-100567), mouse monoclonal anti-Tuj1 (1:500 IF; Fisher Scientific, MAB11905), mouse polyclonal anti-VDAC (1:1000 WB; EMD Millipore, AB10527), rabbit polyclonal anti-SFXN3 (1:50 IF; Sigma, HPA008028).

Techniques: Expressing, Comparison, Fluorescence, Two Tailed Test

Journal: bioRxiv

Article Title: Compartmental Differences in the Retinal Ganglion Cell Mitochondrial Proteome

doi: 10.1101/2024.05.07.593032

Figure Lengend Snippet: Proteomic profiling of mitochondria isolated from RGC somatodendritic and axonal compartments of young (3 mo) and aged (12 mo) mice. A. Representative western blot showing immunoprecipitation of GFP-OMM-tagged mitochondria from RGC somatodendritic and axonal compartments of 3 mo and 12 mo RGC MitoTAG mice compared to control WT mice lacking the tag. Native mitochondrial proteins COX4, and NDUFS4 co-precipitated in RGC MitoTAG but not WT samples. I, input; IP, immunoprecipitate. B. Assigned subcellular localization of proteins identified across two biological replicates in each compartment. C. Volcano plots showing the relative age-related abundance (3mo/12mo) of proteins identified in both age groups. Proteins to the left of the vertical line are more abundant in 12 mo mice and those to the right are more abundant in 3 mo mice. Colored dots represent proteins of interest: SOD2 (purple), SFXN3 (red), and HADHA (blue). D. Comparison of age-related changes in GFP-normalized protein abundance for SOD2, SFXN3, and HADHA for each compartment. Each dot represents an average between two biological replicates.

Article Snippet: The following antibodies with specified dilutions were used for immunofluorescence (IF) and western blot (WB) analyses: goat polyclonal anti-GFP ([1:1000 WB, 1:750 IF]; abcam, ab5450), rabbit polyclonal anti-HADHA (1:50 IF; abcam, ab54477), rabbit polyclonal anti-SOD2/MnSOD (1:50 IF; abcam, ab13534), mouse monoclonal anti-NDUFS4 (1:1000 WB; Santa Cruz Biotechnology, sc-100567), mouse monoclonal anti-Tuj1 (1:500 IF; Fisher Scientific, MAB11905), mouse polyclonal anti-VDAC (1:1000 WB; EMD Millipore, AB10527), rabbit polyclonal anti-SFXN3 (1:50 IF; Sigma, HPA008028).

Techniques: Isolation, Western Blot, Immunoprecipitation, Comparison