Mice cytokine/chemokine magnetic bead panel (MCYTOMAG-70K, Merck Millipore, Darmstadt, Germany) was used for the Milliplex ELISA. The concentrations of 3 human mediators (IL-1β, IL-6, and TNF-α) were measured using a multiplex assay instrument (MAGPIX; Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer’s instructions. Standard curves of known concentrations of recombinant human cytokines (R&D Systems, Minneapolis, MN) were used to convert fluorescence units to cytokine concentration (picograms/milliliter). The protein concentration of each sample was determined using BCA method (G-Biosciences, St. Louis, MO, USA).
Standard curves of known concentrations of recombinant human cytokines
Manufactured by R&D Systems
Standard curves of known concentrations of recombinant human cytokines. This lab equipment provides a visual representation of the relationship between the concentration of specific cytokines and their corresponding signal or response.
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2 protocols using standard curves of known concentrations of recombinant human cytokines
1
Multiplex Cytokine Profiling in Mice
2
Multiplex Analysis of Vitreous Mediators
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Multiplex proteomic analysis was used to measure the levels of inflammatory cytokines, chemokines, and growth factors in the vitreous samples. The MILLIPLEX Human Cytokine/Chemokine/Growth Factor Panel A–Immunology Multiplex Assay (HCYTA-60K-08; MilliporeSigma, Burlington, MA) was used for the MILLIPLEX enzyme-linked immunosorbent assay (ELISA). The concentrations of eight human mediators (IL-1β, IL-1α, IL-6, IL-8, IL-10, IFN-γ, TNF-α, granulocyte-macrophage colony-stimulating factor [GM-CSF]) were measured using a MAGPIX multiplex assay instrument (Bio-Rad Laboratories, Hercules, CA) according to the manufacturer's instructions. Standard curves of known concentrations of recombinant human cytokines (R&D Systems, Minneapolis, MN) were used to convert fluorescence units to cytokine concentration (pg/mL). The protein concentration of each sample was determined using the bicinchoninic acid (BCA) method (G-Biosciences, St. Louis, MO).
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