Interleukin 8 human ELISA kit

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.

Interleukin 10 human ELISA kit

1. INTENDED USE
Immunoenzymetric assay for the in vitro quantitative measurement of human interleukin-10 (IL-10) in serum.

2. CLINICAL BACKGROUND
A. Biological activities
Human interleukin-10 (IL-10) is a 19 kDA lymphokine produced by T helper lymphocytes,by monocytes, macrophages and B-lymphocytes. IL-10 was first characterized as a cytokine synthesis inhibitory factor (CSIF) able to inhibit cytokine synthesis by TH1 clones activated in the presence of antigen presenting cells. However, in the absence of monocytes, IL-10 directly inhibits the growth of T-cells triggered by immobilized anti-CD3 MoAb. This proliferation inhibition was found to be a result of specific inhibition of IL-2 production by the responding T-cells. In vitro, Il-10 is a very powerful inhibitor of monokines (including TNF-α, IL-1, IL-6 and IL-8) produced by LPS-activated monocytes and macrophages. The addition of IL-10 to B lymphocytes results in limited cell proliferation but most importantly in very high immuno- globulin production, a result of the transformation of B-cells into plasma cells. Finally, natural killer (NK) cells appear to be another target for the anti-inflammatory properties of IL-10. Indeed, recent data have shown that IL-10 can inhibit antigen induced IFN-γ production by NK-cells by inhibiting not only production but also the stimulatory effects of IL-12 and TNF on IFN-γ production.

B. Clinical application
So far, circulating levels of IL-10 have been found in serum of patients suffering of Non-Hodgkin's lymphoma, multiple myeloma, cerebral malaria or septic shock.

3. PRINCIPLES OF THE METHOD
The Demeditec IL-10-ELISA is a solid phase Enzyme Amplified Sensitivity Immunoassay performed on microtiterplate. The assay uses monoclonal antibodies (MAbs) directed against distinct epitopes of IL- 10. 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-10 – 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 meas- uring the absorbance, which is proportional to the IL-10 concentration.
A calibration curve is plotted and IL-10 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.