Human SOD3/EC-SOD Antibody

Detection of Human SOD3/EC‑SOD by Western Blot. Western blot shows lysates of human aorta tissue. PVDF Membrane was probed with 0.2 µg/mL of Goat Anti-Human SOD3/EC-SOD Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3420) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for SOD3/EC-SOD at approximately 30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

SOD3/EC‑SOD in Human Kidney. SOD3/EC-SOD was detected in immersion fixed paraffin-embedded sections of human kidney using Goat Anti-Human SOD3/EC-SOD Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3420) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm of epithelial cells in convoluted tubules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human SOD3/EC‑SOD by Simple Western^TM. Simple Western lane view shows lysates of human kidney tissue, loaded at 0.2 mg/mL. A specific band was detected for SOD3/EC-SOD at approximately 38 kDa (as indicated) using 10 µg/mL of Goat Anti-Human SOD3/EC-SOD Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3420) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Mouse SOD3/EC-SOD by Western Blot SOD3-induced VEC expression requires SOD3 enzyme activity and NO. g SOD3 levels in cells as in f; bottom, filter rehybridized with actin. SOD3/actin ratio indicated (n = 3). h FITC-dextran permeability of 1G11-mock and -SOD3 monolayers, untreated or VEGF-pretreated (n = 9). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29422508), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse SOD3/EC-SOD by Immunohistochemistry EC-specific SOD3 expression is sufficient to increase Doxo effect. a Conditional expression allele for the SOD3-IRES-GFP gene at the ROSA26 locus. b SOD3 (green) and CD31 (red) detection in sections from tumors grown in Cre-expressing and Cre− mice after tamoxifen induction. c LLC tumor growth kinetics in SOD3EC-Tg and control (Cre−) mice. Arrows indicate tamoxifen and Doxo treatments (n = 10/group). d Doxo quantification in extracts of tumors dissected at day 16 from control and SOD3EC-Tg mice (n = 15 or 17 mice/group). e, f Images and quantification of lectin-FITC-perfused and CD31-stained vessels in control and SOD3EC-Tg tumors (0.7–1.2 cm3) dissected at day 20. g CD31 staining of control and SOD3EC-Tg tumors at end point. h–l CD31+ structure density (h), mean area covered by CD31+ structures (i), mean vessel length (j) and diameter (k), and the number of vessel branches (l), determined in images as in g. m CD31 (green) and VEC (red) staining of tumors grafted in SOD3EC-Tg and control mice. Bottom panels show red staining in a magnified area. n Quantification of the VEC area in CD31+ structures from images as in m. o Vessel permeability determined by FITC-lectin and Texas Red-dextran in tumors dissected at day 17 from control and SOD3EC-Tg mice. For b, e–o, 10–20 images were analyzed from 5 mice/group; *p < 0.05, **p < 0.01, two-tailed Student’s t-test. Bar, 50 μm Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29422508), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse SOD3/EC-SOD by Immunohistochemistry SOD3 upregulation enhances Doxo chemotherapeutic effects. a, b LLC tumor growth kinetics in Vhcl-, Lov-, Doxo+Vhcl-, or Doxo+Lov-treated WT (a) and SOD3−/− mice (b). Arrows indicate treatment schedule (n = 10 mice/group). c Tumors from WT or SOD3−/− mice treated as above were dissected on day 16 and Doxo was quantified in tumor extracts. d Vhcl- and Lov-treated tumors from WT or SOD3−/− mice were dissected on day 18 (<1 cm3), and SOD3 and CD31 were detected in cryosections by immunohistochemistry (IHC); the two-color merge is shown (n = 10 fields/group; 4 mice/group). e 3-NT detection in paraffin sections of LLC tumors as in d (n = 15 fields/group; 4 mice/group). f LLC-GFP cells were implanted in WT or SOD3−/− mice, Lov-treated as in a, and tumors were dissected on day 21. LLC cells, ECs, and leukocytes were isolated by cell sorting and SOD3 mRNA was determined by qPCR. Data shown as mean ± SEM of triplicates (n = 5 mice/group). g Detection of SOD3 expression (red) and CD31 (green) in sections of Ad-C- or Ad-mSOD3-injected LLC tumors (dissected on day 18); nuclei are DAPI-stained (blue). Arrows in the SOD3 panel indicate the position of CD31+ cells. Bottom panel shows SOD3 fluorescence intensity quantified by ImageJ (15 images/group; 4 mice/group). h Growth kinetics of Ad-C- or Ad-mSOD3-injected LLC tumors. Arrows indicate treatment schedule (n = 9 mice/group). i Doxo quantification in extracts of tumors dissected on day 16 from mice treated as in h. *p < 0.05, **p < 0.01, ***p < 0.001 one-way ANOVA with Dunnett’s post-hoc test using Vhcl group as reference (a, b) or two-tailed Student’s t-test (f–i). Bar, 10 μm (d, g) and 50 μm (e) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29422508), licensed under a CC-BY license. Not internally tested by R&D Systems.