Measurement of IL-8 release into bathing supernatants was performed using the human IL-8 Cytoset (Invitrogen) according to the manufacturer’s recommendations. Briefly, monoclonal antibodies specific for IL-8 were pre-coated onto a MaxiSorp F96 Immunoplate (NUNC, Denmark) and incubated overnight at 4 °C. The following day, the plate was washed with PBS-0.05 % Tween 20 and 100 μl of standards and samples added to each well along with 50 μl of working detection antibody. The plate was incubated for 2 h at room temperature with continual shaking and then washed 5 times before the addition of 100 μl of streptavidin-HRP. The plate was incubated for a further 30 min, washed and the reaction developed with 100 μl of TMB (3,3′,5,5′-tetramethylbenzidine) (KPL, USA). The intensity of the colour was read at both 650 and 450 nm using a Fluostar Omega® microplate reader (BMG Labtech, Germany). Data analysis was performed using BMG Analysis Software (BMG Labtech).
Maxisorp f96 immunoplate
Manufactured by Thermo Fisher Scientific
Sourced in Denmark
The MaxiSorp F96 Immunoplate is a high-quality laboratory equipment designed for various immunoassay applications. It features a flat-bottomed 96-well format and is made of polystyrene material that provides a high-binding capacity surface for efficient capture of biomolecules. The plate is optimized for use in enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs), and other immunological techniques.
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Lab products found in correlation
2 protocols using maxisorp f96 immunoplate
1
Quantifying IL-8 Secretion in Cell Cultures
2
Quantification of IL-8 Secretion
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Measurement of IL-8 release into bathing supernatants was performed using the human IL-8 Cytoset (Invitrogen) according to the manufacturer’s recommendations. Briefly, monoclonal antibodies specific for IL-8 were pre-coated onto a MaxiSorp F96 Immunoplate (NUNC, Denmark) and incubated overnight at 4 °C. The following day, the plate was washed with PBS-0.05 % Tween 20 and 100 μl of standards and samples added to each well along with 50 μl of working detection antibody. The plate was incubated for 2 h at room temperature with continual shaking and then washed 5 times before the addition of 100 μl of streptavidin-HRP. The plate was incubated for a further 30 min, washed and the reaction developed with 100 μl of TMB (3,3′,5,5′-tetra-methylbenzidine) (KPL, USA). The intensity of the colour was read at both 650 and 450 nm using a Fluostar Omega® microplate reader (BMG Labtech, Germany). Data analysis was performed using BMG Analysis Software (BMG Labtech).
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