Cox-2 Antibody Staining Protocol for Immunohistochemistry


Description: Prostaglandin G/H synthase (PGHS) catalyzes the first two steps in prostaglandin, thromboxane and prostacyclin synthesis. Two forms of PGHS have now been identified, cloned and characterized. Both forms of PGHS are heme proteins composed of two 70,000 Dalton subunits, and possess both cyclooxygenase and peroxidase activity. Inhibition of the cyclooxygenase activity of PGHS is responsible for the anti-inflammatory activity of aspirin, indomethacin, ibuprofen, piroxicam, and other non-steroidal anti-inflammatory drugs (NSAIDs). The peroxidase activity of PGHS catalyzes oxidation of a broad range of foreign compounds. Whereas PGHS-1 (COX-1) is constitutively expressed in certain cell types, PGHS-1 and PGHS-2 (COX-2)  are induced in response to a number of hormonal and membrane active agents. This antibody was generated against a synthetic peptide corresponding to a distinct C-terminal region of murine PGHS-2. It specifically reacts with PGHS-2 from most species thus far examined. It does not cross-react with PGHS-1.


Primary Antibody

Name: Cox-2 (PGHS-2) Antibody

Clone: Rabbit anti-Mouse polyclonal

Supplier: Oxford Biomedical

Catalog Number: PG26

Dilution: 1:1000 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 minutes/room temperature

Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95 ºC - 100 ºC /room temperature
Heat/Cool Time: 20 minutes/20 minutes

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Cytoplasmic/peri-nuclear
Images: Search image

Additional Information:
Tissue Type: Colon, kidney
Fixation: Formalin fixed paraffin sections
Positive Control: Colon, kidney
Negative Control: Omit primary antibody, isotype control or absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary


Not tested on frozen sections