Human COX-2 Antibody

Detection of Human COX‑2 by Western Blot. Western blot shows lysates of human peripheral blood mononuclear cells (PBMC) untreated (-) or treated (+) with LPS. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human COX-2 Monoclonal Antibody (Catalog # MAB4198), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for COX-2 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

COX‑2 in A549 Human Cell Line. COX-2 was detected in immersion fixed A549 human lung carcinoma cell line using Mouse Anti-Human COX-2 Monoclonal Antibody (Catalog # MAB4198) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

COX‑2 in Human Breast Cancer Tissue. COX-2 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Mouse Anti-Human COX-2 Monoclonal Antibody (Catalog # MAB4198) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human COX-2 by Western Blot Effect of monocyte-ASC coculture on levels of monocyte subsets, COX-2, PGE2, and EP4. First, monocytes from healthy donors and sepsis patients were cultured alone or cocultured directly with ASCs. The percentage of CD14++CD16+ (a), CD14+CD16++ (b), and CD14++CD16– (c) monocytes in the total monocyte population were determined via flow cytometry. Box and whisker plots represent median (lines within boxes), interquartile range (bounds of boxes), and error bars (upper and lower range); n = 25 for healthy donors and n = 23 for sepsis patients. ***p < 0.001. Then, ASCs and monocytes from sepsis patients were cultured alone, cocultured directly, or cocultured via Transwell for 24 h. Culture supernatants from the above wells were harvested for quantification of PGE2 via ELISA (d). Lysates from different groups were analyzed for COX-2 and EP4 levels via Western blotting. Representative blots and normalized COX-2 levels (e) and EP4 (f) are shown. beta -actin was used as a protein-loading control. Data are expressed as mean ± SEM; n = 5 per group. *p < 0.05, **p < 0.01. ASC adipose-derived mesenchymal stem (stromal) cell, COX-2 cyclooxygenase-2, EP4 prostaglandin E2 receptor 4, MO monocytes, PGE2 prostaglandin E2 Image collected and cropped by CiteAb from the following publication (https://stemcellres.biomedcentral.com/articles/10.1186/s13287-017-0546-x), licensed under a CC-BY license. Not internally tested by R&D Systems.