Cytokine magnetic bead panel

EAE mice were euthanized at peak of the disease (17 days after EAE induction) with an overdose of pentobarbital sodium (Dolethal) and transcardially perfused with 60 mL of sterile saline (0.9% NaCl). Spinal cords were harvested and rapidly frozen in liquid nitrogen. Protein isolation from the spinal cord samples and cytokine quantification were performed as described previously [28 ]. The protein levels of the following cytokines (IL-4, IL-10, IL-1α, IL-1β, IL-3, IL-6, TNF-α, IFN-γ, IL-17A, CSF-3, CCL-5, CCL-2, CXCL-2 and CXCL-10) were analysed using a custom-designed Cytokine Magnetic Bead Panel (Invitrogen) on a MAGPIX system (Millipore).

EAE mice were euthanised 3 days after EAE onset with an overdose of pentobarbital sodium (Dolethal). Blood (500 μl) was obtained by cardiac puncture and centrifuged at 25,000 g for 10 min at room temperature to collect plasma separately from blood cells. Then, mice were transcardially perfused with 60 mL of sterile saline (0.9% NaCl). Spinal cords were harvested and rapidly frozen in liquid nitrogen. Protein isolation from the spinal cord samples and cytokine quantification was performed as we described previously (33 (link)). Protein levels of 6 cytokines (IL-1β, IL-18, IL-6, CXCL-1, TNFα, and IL-10) were analyzed using a custom-designed Cytokine Magnetic Bead Panel (Invitrogen) on a MAGPIX system (Millipore).