Human/Mouse COX-2 Antibody AF4198: R&D Systems

Human/Mouse COX-2 Antibody

(1 Review)
  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human and mouse COX-2 in Western blots. In Western blots, less than 1% cross-reactivity with recombinant human COX-1 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human COX-2
    Ala18-Leu604
    Accession # P35354
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunocytochemistry
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.
Immunocytochemistry
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.
Intracellular Staining by Flow Cytometry
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
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    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.
Background: COX-2

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.

  • Long Name:
    Cyclooxygenase 2
  • Entrez Gene IDs:
    5743 (Human); 19225 (Mouse); 29527 (Rat)
  • Alternate Names:
    COX2; COX-2; COX2cyclooxygenase 2b; cyclooxygenase-2; EC 1.14.99; EC 1.14.99.1; GRIPGHS; hCox-2; PGG/HS; PGH synthase 2; PGHS-2; PHS II; PHS-2; PHS-II; prostaglandin G/H synthase 2; prostaglandin G/H synthase and cyclooxygenase; Prostaglandin H2 synthase 2; prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase andcyclooxygenase); Prostaglandin-endoperoxide synthase 2; PTGS2
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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 species: Mouse.
We have 1 review tested in 1 application: Immunocytochemistry/Immunofluorescence.

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Images Ratings Applications Species Reviewed By Date Details
Immunocytochemistry/Immunofluorescence Human/Mouse COX-2 Antibody AF4198
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Excellent
 ICC/IF Mouse Anonymous 05/15/2017
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Immunocytochemistry/Immunofluorescence Human/Mouse COX-2 Antibody AF4198
Immunocytochemistry/Immunofluorescence: Human/Mouse COX-2 Antibody [AF4198]

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedSpleen tissue
SpeciesMouse

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