Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total Cox2 Sandwich ELISA Kit #7291

When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

Kit Includes Volume Solution Color
Cox2 Mouse Ab Coated Microwells 96 tests
Cox2 Rabbit Detection Ab 11 ml Green
Anti-Rabbit IgG, HRP-linked Antibody 11 ml Red
TMB Substrate #7004 11 ml Colorless
STOP Solution #7002 11 ml Colorless
Sealing Tape 2 sheets
Cell Lysis Buffer (10X) #9803 15 ml Yellowish
ELISA Sample Diluent 25 ml Blue
ELISA Wash Buffer (20X) 25 ml Colorless

Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Description

The PathScan® Total Cox2 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Cox2. A Cox2 Mouse Antibody has been coated onto the microwells. After incubation with cell lysates, Cox2 protein is captured by the coated antibody. Following extensive washing, a Cox2 Rabbit Detection Antibody is added to detect the captured Cox2 protein. Anti-rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of Cox2.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

PathScan® Total Cox2 Sandwich ELISA Kit detects endogenous levels of total Cox2 protein. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

ELISA

ELISA

Figure 2. The relationship between the protein concentration of lysates from CCD-1070Sk cells, serum starved or hIL-1β-treated, and the absorbance at 450 nm is shown. CCD-1070Sk cells (80-90% confluent) were serum starved or treated with hIL-1β #8900 (5 ng/ml, 16 hr) and then lysed.

ELISA - Western correlation

ELISA - Western correlation

Figure 1. Treatment of CCD-1070Sk cells with IL-1β stimulates expression of Cox2 as detected by the PathScan® Total Cox2 Sandwich ELISA Kit. CCD-1070Sk cells (80-90% confluent) were serum starved or treated with hIL-1β #8900 (5 ng/ml, 16 hr). The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blot using Cox2 Antibody #4842 is shown in the bottom figure.

Background

Cyclooxygenase1 (Cox1) and cyclooxygenase2 (Cox2), family members with 60% homology in humans, catalyze prostaglandin production from arachidonic acid (1,2). While Cox1 expression is constitutive in most tissues, Cox2 expression is induced by lipopolysaccharide (LPS) and peptidoglycan (PGN) (3). PGN activates Ras, leading to phosphorylation of Raf at Ser338 and Erk1/2 at Tyr204. The activation of MAP kinase signaling results in subsequent activation of IKKα/β, phosphorylation of IκBα at Ser32/36, and NF-κB activation. Finally, activation of the transcription factor NF-κB is responsible for the induction of Cox2 expression (4). Investigators have shown that LPS and PGN induce the clinical manifestations of arthritis and bacterial infections, such as inflammation, fever, and septic shock (5). Research studies have indicated that Cox1 and Cox2 may also play a role in the neuropathology of Alzheimer's disease by potentiating γ-secretase activity and β-amyloid generation (6).

  1. Xie, W.L. et al. (1991) Proc Natl Acad Sci USA 88, 2692-6.
  2. Vane, J.R. et al. (1998) Annu Rev Pharmacol Toxicol 38, 97-120.
  3. O'Neill, G.P. et al. (1994) Mol Pharmacol 45, 245-54.
  4. Chen, B.C. et al. (2004) J Biol Chem 279, 20889-97.
  5. Wang, Q. et al. (2001) Infect Immun 69, 2270-6.
  6. Qin, W. et al. (2003) J Biol Chem 278, 50970-7.

Application References

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Protocols

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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