Cyclooxygenase (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.
Human/Mouse Total COX-2 DuoSet IC ELISA
R&D Systems | Catalog # DYC4198-2
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Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
156-10000 pg/mL
Sample Type
Cell Lysates
Reactivity
Human, Mouse
Human/Mouse Total COX-2 DuoSet IC ELISA Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Available in 2, 5, and 15- (96-well) plate pack sizes
- Economical alternative to Western blot
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Product Summary for Human/Mouse Total COX-2 DuoSet IC ELISA
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.
Product Specifications
Assay Format
96-well strip plate (sold separately)
Sample Volume Required
Cell lysates (100 µL)
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Scientific Data Images for Human/Mouse Total COX-2 DuoSet IC ELISA
Standard Curve
The Human/Mouse Total COX-2 DuoSet IC ELISA specifically recognizes total COX-2, as shown by Western blot.
Lysate prepared from CCD-1070Sk human foreskin fibroblasts treated with 5.0 ng/mL of recombinant human (rh) IL-1 beta (Catalog # 201-LB) for 16 hours was incubated in wells coated with Human/Mouse Total COX-2 Capture Antibody. Unbound material was removed by washing and bound material was solubilized in SDS gel sample buffer. The same lysate and the captured protein were electrophoresed, transferred to a PVDF membrane, and immunoblotted with an Anti-COX-2 Polyclonal Antibody (Catalog # AF4198). Only the band corresponding to human/mouse COX-2 was detected in captured material.
Quantification of induced COX-2.
Human PBMCs, THP-1 human acute monocytic leukemia cells, and RAW 264.7 mouse monocyte/macrophage cells were cultured in the absence or presence of 100 nM Phorbol 12-myristate 13-acetate (PMA) for 48 hours and 1.0 µg/mL of Lipopolysaccharide (LPS) for an additional 24 hours. Human CCD-1070Sk cells were cultured in the absence or presence of 5.0 ng/mL of rhIL-1 beta for 16 hours. The lysates were quantified with this DuoSet IC ELISA. The same lysates were immunoblotted (inset) with an anti-COX-2 polyclonal antibody. The DuoSet IC ELISA results correlate well with the relative amounts of human/mouse COX-2 detected by Western blot.Kit Contents for Human/Mouse Total COX-2 DuoSet IC ELISA
- Capture Antibody
- Conjugated Detection Antibody
- Calibrated Immunoassay Standard or Control
- Streptavidin-HRP
Other Reagents Required
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Lysis Buffer*
IC Diluent*
Blocking Buffer*
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent
*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product
Background: COX-2
Long Name
Cyclooxygenase 2
Alternate Names
COX2, PGHS-2, PHS-II, PTGS2
Gene Symbol
PTGS2
Additional COX-2 Products
Product Documents for Human/Mouse Total COX-2 DuoSet IC ELISA
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human/Mouse Total COX-2 DuoSet IC ELISA
For research use only
Citations for Human/Mouse Total COX-2 DuoSet IC ELISA
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
- How to Run an R&D Systems DuoSet ELISA
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- Quantikine HS ELISA Kit Assay Principle, Alkaline Phosphatase
- Quantikine HS ELISA Kit Principle, Streptavidin-HRP Polymer
- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- Troubleshooting Guide: ELISA
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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