The Human Cytokine Array (ARY005) kits were purchased and used to simultaneously detect relative expression levels of 36 human cytokines (R&D Systems, Oxon, UK). Conditioned medias from each sample, 0.5 ml respectively, were mixed with a cocktail of biotinylated detection antibodies. The mixture was then incubated with the Human Cytokine Array Panel A membrane. A wash step was needed to remove unbound material, and then Streptavidin-HRP and chemiluminescent detection reagents were added. Light was produced at each spot in proportion to the amount of analyte bound. The positive signals on developed films were identified by placing the transparency overlay on the ChemiDoc imaging system (Bio-Rad) and aligning it with the reference spots in three corners of each array. Pixel density was analyzed with ImageJ (Wayne Rasband, NIH).
Human cytokine array ary005 kits
Manufactured by R&D Systems
Sourced in United Kingdom
The Human Cytokine Array (ARY005) kits from R&D Systems are designed to detect and quantify the relative levels of multiple human cytokines and chemokines simultaneously in a variety of sample types. The kits use a membrane-based antibody array format to capture and detect target analytes in a single experiment.
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2 protocols using human cytokine array ary005 kits
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Profiling Cytokine Expression by Array
2
Profiling Cytokine Expression by Array
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The Human Cytokine Array (ARY005) kits were purchased and used to simultaneously detect relative expression levels of 36 human cytokines (R&D Systems, Oxon, UK). Conditioned medias from each sample, 0.5 ml respectively, were mixed with a cocktail of biotinylated detection antibodies. The mixture was then incubated with the Human Cytokine Array Panel A membrane. A wash step was needed to remove unbound material, and then Streptavidin-HRP and chemiluminescent detection reagents were added. Light was produced at each spot in proportion to the amount of analyte bound. The positive signals on developed films were identified by placing the transparency overlay on the ChemiDoc imaging system (Bio-Rad) and aligning it with the reference spots in three corners of each array. Pixel density was analyzed with ImageJ (Wayne Rasband, NIH).
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