Human/Mouse Total COX-2 DuoSet IC ELISA Summary Assay Type | Solid Phase Sandwich ELISA | Format | 96-well strip plate | Assay Length | 4 hours 40 mins (after plate preparation) | Sample Type & Volume Required Per Well | Cell lysates (100 μL) | Sensitivity | 5 pg/mL | Assay Range | 31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma) | Specificity | 156.0 - 10,000 pg/mL | Cross-reactivity | < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested | Interference | No significant interference observed with available related molecules. |
Product Summary This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total COX-2 in cell lysates. An immobilized capture antibody specific for COX-2 binds both phosphorylated and unphosphorylated COX-2. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format. Preparation and Storage Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. | Storage | Store the unopened product at 2 - 8 °C. Do not use past expiration date. |
Background: COX-2Cyclooxygenase (COX), also known as prostaglandin G/H synthase, is a membrane bound enzyme partly responsible for the catalysis of prostanoid synthesis. COX is expressed as at least three different isoforms. COX-1 is constitutively expressed and thought to regulate a number of 'housekeeping' functions such as vascular hemostasis, renal blood flow, and glomerular function. COX-2 expression is tightly regulated and induced by inflammatory mediators such as growth factors, cytokines, and endotoxin. COX-3 appears to be much more strictly regulated spatially and is observed in greatest abundance in cerebral cortex. 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 |
Assay Procedure Refer to the product datasheet for the complete assay procedure. Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate. 1. Prepare all reagents, standard dilutions, and samples as directed in the product insert. 2. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal. 3. Add 50 μL of Assay Diluent to each well. 4. Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours. 5. Aspirate each well and wash, repeating the process 4 times for a total of 5 washes. 6. Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours. 7. Aspirate and wash 5 times. 8. Add 100 μL Substrate Solution to each well. 9. Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm. |
