COX-2

Cyclooxygenase (COX) is an enzyme that is responsible for formation of important biological mediators called prostanoids (including prostaglandins, prostacyclin and thromboxane). Pharmacological inhibition of COX can provide relief from the symptoms of inflammation and pain; this is the method of action of well-known drugs such as aspirin and ibuprofen. COX-2 inhibition by nonsteroidal anti-inflammatory agents has been shown to decrease angiogenesis and tumor growth, and promote apoptosis.

COX-2 overexpression has been associated with increased microvascular density, and VEGF protein expression in head and neck Squamous-cell Carcinomas and is a poor prognostic indicator in this entity as well. COX-2 overexpression has also been suggested as a poor prognostic indicator in Carcinomas of the colon, breast, pancreas, and Adenocarcinomas of the lung.

Anti-COX-2 is a rabbit monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.

1. Stoehlmacker J, Lenz Heinz-Josef, Semin Oncol 30.(3)suppl6(June),2003:10-16
2. Gallo O, et al. Hum Pathol. 33:708-714
3. Sano H, et al. Cancer Res. 1995;Sep 1;55(17):3785-9
4. Denkert C, et al. Cancer. 2003;Jun 15;97(12):2978-87
5. Sheehan KM, et al. Hum Pathol. 34:1242-1246

1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
2. Air dry for 2 hours at 58° C.
3. Deparaffinize, dehydrate and rehydrate tissues.
4. Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
5. Any of three heating methods may be used:
   • Electric Pressure Cooker
     Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature.
   • Water Bath Method
     Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes.
   • Conventional Steamer Method
     Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
7. Wash slides with IHC wash buffer or DI water.
8. Continue IHC staining protocol.