The cytokine content of the culture medium was assessed via Proteome Profiler Human Cytokine Array (Panel A Kit, R&D Systems, Minneapolis, USA). Culture medium from neutrophils treated with or without nanoparticles (0.75 mL) was mixed with 0.75 mL of Array Buffer 4. A cocktail of biotinylated antibodies was added and incubated at room temperature for 1 hr. The sample-antibody mixture was added to the blocked membrane and incubated at 4 °C for 18 hr. Following 3 × 10 min washes, streptavidin-HRP antibody was added at a dilution of 1:2000 and incubated at room temperature for 45 min. The membranes were washed, probed with ECL Western Blotting Substrate (Thermo Scientific) and imaged using a ChemiDoc™ XRS + System (BioRad). The intensity of each spot was measured using the ImageJ software and the average pixel density of each duplicate spot was expressed for comparative analysis.
Proteome profiler human cytokine array panel a kit
Manufactured by R&D Systems
Sourced in United States
The Proteome Profiler Human Cytokine Array (Panel A Kit) is a multiplex array designed for the simultaneous detection and relative quantification of 36 different human cytokines, chemokines, and other soluble proteins in a single experiment. The kit includes a membrane-based antibody array, detection antibodies, and all necessary reagents.
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Lab products found in correlation
2 protocols using proteome profiler human cytokine array panel a kit
1
Cytokine Profiling of Nanoparticle-Treated Cells
2
Plasma Cytokine Profiling by Antibody Array
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The relative concentration of plasma cytokine protein was determined by cytokine array and was assessed using the Proteome Profiler Human Cytokine Array (Panel A Kit, R&D Systems, Minneapolis, MN, USA). Arrays were used according to manufacturer's instructions. Briefly, plasma was incubated with nitrocellulose membrane coated with 36 different capture antibodies. Bound cytokines were detected with a cocktail of secondary biotinylated antibodies and streptavidin-HRP. Membranes were imaged using ChemiDocT-MXRS + System (Bio-Rad, Hercules, CA, USA). ImageJ software (Version 1.5.1, 2018) was used to determine the average pixel density of the duplicate spots.
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