INTERLEUKIN 8 HUMAN ELISA

BRAND:                 DEMEDITEC DIAGNOSTICS GMBH

PRODUCT CODE: DE4700

AVAILABILITY:     IN STOCK

DESCRIPTION

• CE certified for IVD use

• 96 well ELISA microplate

• Incubation time: 2 h 30 min

• Range: 40.2 - 1,845 pg/ml

• Sensitivity: 1.1 pg/ml

• Sample size: 100 µl

• Sample type: plasma

• Substrate: TMB 450nm

1. INTENDED USE

Immunoenzymetric assay for the in vitro quantitative measurement of human interleukin-8 (IL-8) in plasma.

2. CLINICAL BACKGROUND

A. Biological activities

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 proinflamma- tory 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.

B. Clinical application

The IL-8 level in the septic shock patients was found to correlate with mortality and in acute graft liver rejection the IL-8 serum levels were reported to have markedly increased. The level of IL-8 in these or other conditions may prove to be important in characterizing the progress of these disease conditions.

3. PRINCIPLES OF THE METHOD

The Demeditec IL-8-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 incu- bated. 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 meas- uring 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.