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.
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 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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Cyclooxygenase-2 (COX-2) also known as prostaglandin G/H synthase 2 (PGHS2) is a 70 kDa microsomal enzyme that belongs to the prostaglandin G/H synthase family. It is inducibly-expressed by a number of cell types, including fibroblasts, vascular smooth muscle cells, endothelium, and monocytes. Functionally, COX-2 is a homodimer that catalyzes two steps in the conversion of arachadonic acid to prostaglandin H2. Mature human COX-2 is 587 amino acids (aa) in length and contains one EGF-like domain (aa 18‑55), a potential membrane interacting region (aa 277‑292) and a globular catalytic domain (aa 293‑604). At least one splice form exists that shows an 11 aa substitution for the C-terminal 451 amino acids. Mature human COX-2 shows 87% aa identity to mouse COX-2.
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