CBA053

COX-2 ELISA Kit

• Synonyma: Cyclooxygenase-2 ELISA Kit
• UNSPSC Code: 41116133

Vlastnosti

• usage: sufficient for 96 tests
• species reactivity: human
• packaging: pkg of 1 96-well plate(s)
• manufacturer/tradename: Calbiochem^®
• storage condition: OK to freeze
• assay range: standard curve range: 2.5-200 ng/mL
• input: sample type cell lysate; sample type tissue extract(s)
• detection method: colorimetric
• storage temp.: −20°C

Popis

General description

This product has been discontinued.

This kit detects and quantifies human COX-2 in cell lysates and tissue extracts. Cyclooxygenase (COX) enzymes catalyze the synthesis of prostaglandins from arachidonic acid. COX-2 expression induced by various inflammatory and/or mitogenic stimuli such as cytokines, growth factors or tumor promoters.

Biochem/physiol Actions

The Calbiochem COX-2 ELISA Kit is intended for the quantitative measurement of COX-2 in human cell lysates. Serum and plasma samples are not recommended due to inherently low concentrations of COX-2 in biological fluids.

Preparation Note

Cell lysates: Most mammalian cells do not express significant amounts of COX-2, so in order to detect COX-2 in lysates of most cell types; COX-2 must be induced by an appropriate stimulus. Below is a sample protocol for COX-2 induction in U937 cells:1. Prepare all reagents immediately prior to use. 2. Grow U937 cells in complete RPMI 1640 medium.3. Harvest the cells during the log phase of growth.4. Adjust the cell density to 1 x 10⁶ cells/ml and incubate for 3 days in the presence of 15 nM PMA.5. Harvest the cells and incubate for 24 h in fresh complete RPMI 1640 medium.6. Stimulate the cells with LPS for 6 h at a concentration of 250 ng/ml.7. Lyse the cells with CytoBuster Protein Extraction Reagent (Cat. No. 71009) according to the recommended protocol.8. Determine the protein concentration. A concentration of 2-5 mg/ml is recommended.9. Store the cell lysate at -70°C until use.

Note: Allow all kit components and samples to reach room temperature (18-25°C) prior to assay. Prepare all reagents immediately prior to use. The following table provides reagent preparation instructions to obtain a volume of reagent sufficient for one 2x8-well strip (16 wells).

Table 1: Reagent Preparation

4. Standards:a. Reconstitute the COX-2 Standard with 1000 µl deionized or distilled water to yield a stock solution of 2000 ng/ml. Dispense unused reconstituted COX-2 Standard into aliquots and freeze (-70°C). Aliquots are stable for several months at -70°C.b. Prepare a standard curve by making dilutions of the reconstituted COX-2 Standard in Assay Diluent as follows:

Table 2: Sample Dilutions

Upon arrival store the entire contents of the kit at -20°C.

Analysis Note

Positive Control
U937 cells

Other Notes

Anti-COX-2 Coated 96-Well Plate, COX-2 Standard, COX-2 Detector Antibody, HRP Conjugate, Assay Diluent, ELISA 20X Plate Wash Concentrate, TMB Substrate, ELISA Stop Solution, Plate Sealers, and a user protocol

Blanke, S.R., et al. 2005. Semin. Oncol.32, 69.
Dannenberg, A.J., et al. 2005. J. Clin. Oncol.23, 254.
Sanborn, R. and Blake, C.D., 2005. Semin. Oncol.32, 69.
Kim, H.J., et al. 2003. Int. J. Oncol.22, 99.
Shigemasa, K., et al. 2003. Int. J. Oncol.22, 99.
El-Bayoumy, K., et al. 2002. Int. J. Oncol.20, 557.
Hwang, D., et al. 1998. J. Natl. Cancer Inst.90, 455.

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Toxicity: Multiple Toxicity Values, refer to MSDS (O)