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Proteome profiler mouse cytokine array panel a array kit

Manufactured by R&D Systems

The Proteome Profiler Mouse Cytokine Array Panel A Array Kit is a multiplex ELISA-based array designed to detect the relative levels of 40 different mouse cytokines and chemokines simultaneously in a single sample.

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2 protocols using proteome profiler mouse cytokine array panel a array kit

1

Cytokine Profiling in Immortalized BMDMs

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To measure cytokine production we used the Proteome Profiler Mouse Cytokine Array Panel A Array Kit (R&D Systems Inc.), which consists of individual nitrocellulose membranes each containing 40 capture antibodies spotted in duplicates and simultaneously profiles the relative levels of multiple cytokines in the samples. Briefly, immortalized BMDMs-WT were seeded overnight at 37°C in 5% CO2 in 6-well plates (1×106 cells/well) in a final volume of 2 ml per well. Then, cells were primed with IFN-γ (12.5 U/ml) for 2 hours, then left untreated (medium alone) or treated with nHZ, and incubated for further 24 h. Then, cell supernatants were collected, centrifuged, and then mixed with a cocktail of biotinylated detection antibodies, and subjected to the cytokine array. The streptavidin-HRP and chemiluminescent detection reagents were added sequentially, and the secreted cytokines were detected. The films were scanned (Camag Reprostar 3) and the spot area was determined using densitometric analysis software.
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2

Cytokine and Chemokine Profiling by ELISA and Proteome Array

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Culture supernatants or diluted peritoneal exudates were used for cytokine and chemokine assays by sandwich ELISA, by using pairs of specific mAbs (R&D Systems or eBioscience), one of which was labeled with biotin, and developed with streptavidin-alkaline phosphatase (Invitrogen Life Technologies) and with p-nitrophenyl phosphate (Thermo Scientific Pierce, Waltham, MA, USA) as substrate. Cytokine levels were also analyzed in fresh supernatants by chemiluminescence arrays (Proteome Profiler – Mouse Cytokine Array Panel A Array Kit, R&D Systems), according to the manufacturer's instructions.
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