On-chip immunoassay was performed using the Bio-Plex Pro Human Cytokine IL-6 antibody set, Cytokine Reagent Kit, and cytokine standards (Bio-Rad Laboratories Inc, Hercules, CA, 171304070M, 171D50001, and 171B5006M, respectively). All reagents were prepared as per manufacturer recommendations, with the standards reconstituted and diluted in 0.5% BSA in PBS. The device was operated similarly to the validation experiments described previously with some minor changes. First, Bio-Plex Pro assay buffer was used in the channel blocking and bead collection steps. In addition, Bio-plex Pro wash buffer was used instead of PBS for all wash steps. Finally, the flow rate for the sample, biotinylated detection antibody, and streptavidin-phycoerythrin incubation steps were 5, 50, and 50 nL/min and performed for 4.5, 1.5, and 0.5 hours, respectively. At the completion of the assay, the beads were collected and transferred to a 96 well plate and analyzed using the Bio-Plex 200 system as per manufacturer recommendations. The raw event data for each bead was exported into Matlab for analysis.
Cytokine reagent kit
Manufactured by Bio-Rad
Sourced in United States
The Cytokine Reagent Kit is a laboratory product designed for the detection and quantification of various cytokines. It provides the necessary reagents and components to perform cytokine-related assays and analyses.
Automatically generated - may contain errors
3 protocols using cytokine reagent kit
1
On-Chip Cytokine Immunoassay Optimization
2
Cytokine Dynamics in Mouse Serum
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Cytokines, chemokines, and growth factor levels in mouse serum were quantified at different time points (blood sample time line: before ligation and at 8 h, 24 h, 72 h, 7 days, 14 days, and 21 days after ligation). Serum was stored after a spin‐down procedure at −80°C, and mice were injected with 250 μL saline i.p. after each blood withdrawal. Cytokine concentrations of IL‐1α, IL‐1ß, IL‐2, IL‐3, IL‐4, IL‐5, IL‐6, IL‐9, IL‐10, IL‐12 (p40), IL‐12 (p70), IL‐13, IL‐17, Eotaxin, G‐CSF, GM‐CSF, IFN‐γ, KC, MCP‐1 (MCAF), MIP‐1α, MIP‐1ß, RANTES, TNF‐α, FGF basic, IL‐15, IL‐18, LIF, MCSF, MIG, MIP‐2, PDGF‐BB, and VEGF were determined using the multiplex pro‐mouse cytokine 23‐plex and 9‐plex panels and the cytokine reagent kit (Bio‐Rad Laboratories) as described previously.25 (link) Samples were analyzed as triplets and standards in duplicates using a Luminex‐100 instrument with Bio‐Plex Manager 4.1 software (Bio‐Rad Laboratories).28 (link)
3
Cytokine Profiling in Plasma and Saliva
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Plasma and saliva samples were assayed for cytokine concentrations using multiplexed (IL-1β, IL-4, IL-5, IL-6, IFNγ, and TNFα) bead-based immunoassay kits combined with a Cytokine Reagent Kit (Bio-Rad Laboratories, Hercules, CA, USA) and a Bio-Plex™ Diluent Kit (Bio-Rad Laboratories) in the Bio-Plex™ MAGPIX System (Bio-Rad Laboratories) according to the manufacturer’s protocol. Concentrations of plasma cytokines were calculated using Bio-Plex™ Manager MP Software (Bio-Rad Laboratories).
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