Interleukin-8 human ELISA

>Background:IL-8 (also known as NAP-1 for Neutrophil-activating peptide) is a chemoattractant protein for neutrophils. This cytokine belongs to a new family of chemotactic peptides called "chemokines". This proinflammatory mediator is secreted by different cells such as monocytes, neutrophils, endothelial cells, fibroblast after activation, and by mitogen-stimulated T lymphocytes. IL-8 is a key cytokine that has been found in scales of psoriasis patients, in synovial fluid of patients suffering from rheumatoid arthritis and gout. The role of IL-8 in the recruitment of neutrophils in the lung during ARDS has also been suggested.

Description:
The Interleukin-8 human ELISA is a solid phase Enzyme Amplified Sensitivity Immunoassay performed on microtiterplate. The assay uses monoclonal antibodies (MAbs) directed against distinct epitopes of IL-8. Calibrators and samples react with the capture monoclonal antibody (MAb 1) coated on microtiter well and with a monoclonal antibody (MAb 2) labelled with horseradish peroxidase (HRP). After an incubation period allowing the formation of a sandwich: coated MAb 1 – human IL-8 – MAb 2 – HRP, the microtiterplate is washed to remove unbound enzyme labelled antibody. Bound enzyme-labelled antibody is measured through a chromogenic reaction. Chromogenic solution (TMB) is added and incubated. The reaction is stopped with the addition of Stop Solution and the microtiterplate is then read at the appropriate wavelength. The amount of substrate turnover is determined colourimetrically by measuring the absorbance, which is proportional to the IL-8 concentration. A calibration curve is plotted and IL-8 concentration in samples is determined by interpolation from the calibration curve. The use of the ELISA reader (linearity up to 3 OD units) and a sophisticated data reduction method (polychromatic data reduction) result in a high sensitivity in the low range and in an extended calibration range.

Product features:
The kit contains reagents for 96 determinations;
Microtiter plate consisting of 12x8 (breakapart);
Microtiter plate reader at 450, 490 and 650 nm;
Limit of detection: 1,1 pg/ml;
IVD.