Human/Mouse COX-2 Antibody

Detection of Human and Mouse COX‑2 by Western Blot. Western blot shows lysates of human peripheral blood mononuclear cell (PBMC) and RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with 1 ug/mL LPS for 24 hours and U937 human histiocytic lymphoma cell line untreated or treated with 100 nM PMA and 1 ug/mL LPS for 48 hours and 24 hours, respectively. PVDF membrane was probed with 1 µg/mL of Human/Mouse COX-2 Polyclonal Antibody (Catalog # AF4198), followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). 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 HUVEC Human Cells. COX-2 was detected in immersion fixed HUVEC human umbilical vein endothelial cells using 10 µg/mL Human/Mouse COX-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4198) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

COX‑2 in RAW 264.7 Mouse Cells. COX-2 was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cells stimulated with LPS using Goat Anti-Human/Mouse COX-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4198) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of COX‑2 in RAW 264.7 Mouse Cell Line by Flow Cytometry. RAW 264.7 mouse monocyte/macrophage cell line treated with 1 µg/mL LPS for 24 hours was stained with Goat Anti-Human/Mouse COX-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4198, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Detection of Mouse COX-2 by Western Blot Fibrotic deposit and cardiac dysfunction correlate with decreasing CVPC presence in mdx heart. (A) Representative images of histological analysis stained using Masson trichrome technique showing myocytes (in red) and collagenous fibrotic tissue (in blue) in the left ventricle of WT and mdx hearts at 9, 24, and 52 wo. Line represents 100 µm. (B) The ratio of red and blue stained tissue was evaluated in WT hearts (open bars and black dots, n = 4–11 slices/3 animals per group) and mdx hearts (black bars and grey dots, n = 3–16 slices/3 animals per group) at the age of 24 wo and further at 52 wo. Statistical significance was calculated by Kruskal–Wallis test and Dunn‘s multiple comparison post-hoc test (*** p < 0.001, **** p < 0.0001). (C) Western blot analysis of collagen proteins and inflammatory proteins in the cardiac tissues. Left panel shows representative images of collagen 1A1 (Coll 1A1), collagen 3 (Coll 3), cyclooxygenase 2 (COX-2), and matrix metalloproteinase 9 (MMP-9) compared to the GAPDH control. The right panels show the normalized densitometry of each protein normalized by GAPDH content of WT (open bars, n = 2 animals) and mdx (black bars, n = 2 animals). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/34068508), licensed under a CC-BY license. Not internally tested by R&D Systems.