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Multiparameter flow cytometric analysis of COX-2 expression in activated human CD14+ monocytes. Human peripheral blood mononuclear cells were either cultured alone (Untreated; Left Panel) or cultured (LPS-Treated; Right Panel) for 5 hours in the presence of lipopolysaccharide (LPS; 1 µg/ml; Sigma, Cat. No. L2654). Cells in suspension and adherent cells (collected using BD™ Accutase™ Cell Detachment Solution; Cat. No. 561527) were harvested, washed, and stained with PerCP-Cy™5.5 Mouse Anti-Human CD14 antibody (Cat. No. 562692). The cells were then washed, fixed and permeabilized using the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714) and stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680) or PE Mouse Anti-Human COX-2 antibody (Cat. No. 565125), as indicated. Two-parameter flow cytometric dot plots showing the correlated expression of forward scattered-light signals versus COX-2 (or Ig Isotype control staining) were derived from CD14-positive gated events with the forward and side scattered-light characteristics of intact monocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
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BD Pharmingen™ PE Mouse Anti-Human COX-2
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Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Accutase is a registered trademark of Innovative Cell Technologies, Inc.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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The AS67 monoclonal antibody specifically binds to Cyclooxygenase-2 (COX-2/COX2) which is also known as, Prostaglandin G/H synthase 2 (PGHS-2). The COX-2 enzyme is encoded by the PTGS2 gene, Prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase) and exists as a homodimer that is located on the lumenal surface of the endoplasmic reticulum and on the inner and outer membranes of the nuclear envelope. COX-2 catalyzes the conversion of arachidonic acid into prostaglandin H2 (PGH2). Specific prostaglandin synthases then convert the PGH2 intermediate into various biologically active prostanoids, including Prostaglandin E2 (PGE2) and Thromboxane A2 (TXA2). COX-2 expression is not detected in most tissues under physiological conditions. However, COX-2 expression is rapidly upregulated during the course of inflammation, following cellular stresses, and in response to growth factors, tumor promoters, hormones, bacterial endotoxins, and inflammatory cytokines, eg, Interleukin-1α (IL-1α). COX-2 can be induced in a number of cell types, including fibroblasts, endothelial cells, monocytes, ovarian follicles and mesenchymal stromal cells. Although COX-2 is structurally homologous to Cyclooxygenase-1 (COX-1), the AS67 antibody does not crossreact with recombinant human COX-1 protein as tested by immunoassay.

Development References (6)
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Chen EP; Markosyan N; Connolly E et al. Myeloid Cell COX-2 deletion reduces mammary tumor growth through enhanced cytotoxic T-lymphocyte function. Carcinogenesis. 2014; 35(8):1788-1897. (Biology). View Reference
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Fogel-Petrovic M, Long JA, Knight DA, Thompson PJ, Upham JW. Activated human dendritic cells express inducible cyclo-oxygenase and synthesize prostaglandin E2 but not prostaglandin D2. Immunol Cell Biol. 2004; 82(1):47-54. (Biology). View Reference
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Herschman HR. Prostaglandin synthase 2. Biochim Biophys Acta. 1996; 1299(1):125-140. (Biology). View Reference
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Maloney CG; Kutchera WA; Albertine KH; McIntyre TM; Prescott SM; Zimmerman GA. Inflammatory agonists induce cyclooxygenase type 2 expression by human neutrophils. J Immunol. 1998; 160(3):1402-1410. (Biology). View Reference
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Ruitenberg JJ; Waters CA. A rapid flow cytometric method for the detection of intracellular cyclooxygenases in human whole blood monocytes and a COX-2 inducible human cell line. J Immunol Methods. 2003; 274(1-2):93-104. (Clone-specific: Cytometric Bead Array, Flow cytometry, Inhibition). View Reference
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Yu KR, Lee JY, Kim HS, et al. A p38 MAPK-mediated alteration of COX-2/PGE2 regulates immunomodulatory properties in human mesenchymal stem cell aging. PLoS ONE. 2014; 9(8):e102426. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.