Cytokine bead assay was performed with the FlowCytomix System (eBioscience, Frankfurt, Germany) according to the manufacturer’s manual. One half of a brain per mouse was lysed in 50 mM Tris pH 7.4 with cOmplete Ultra Tablet (Roche, Basel, Switzerland) with an Ultra-Turrax (5 section at 17500 rpm) on ice. Lysates were centrifuged at 10000 x g, 4°C for 30 min. Protein levels in supernatants were measured by BCA Protein Assay (Pierce, Rockford, USA).
Flowcytomix system
Manufactured by Thermo Fisher Scientific
Sourced in Germany
The FlowCytomix System is a flow cytometry-based platform designed for multiplex analysis of various analytes, such as proteins, cytokines, and other biomolecules. The system utilizes fluorescent-encoded beads and a flow cytometer to detect and quantify multiple targets simultaneously in a single sample.
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Lab products found in correlation
3 protocols using flowcytomix system
1
Cytokine Profiling in Mouse Brain
2
Cytokine Profiling from Cardiac Puncture
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For cytokine analysis blood was taken by cardiac puncture and serum was isolated via centrifugation at 10.000g for 1 minute. Cytokine concentrations were determined using the FlowCytomix system (formerly ebioscience, now Thermo Fisher Scientific, distributed by Fisher Scientific Austria GmbH, Vienna, Austria). In some experiments, IFNγ concentrations were measured by ELISA (formerly ebioscience, now Thermo Fisher Scientific, distributed by Fisher Scientific Austria GmbH, Vienna, Austria).
3
Macrophage Polarization and Modulation
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Macrophages (M1 and M2) were treated with LPS (Sigma-Aldrich) at 10ng/ml for 24 hours, or pre-treated with ibuprofen or compounds at 5 μM for 30 minutes and after treated with LPS for 24 hours; or pre-treated with LPS for 30 minutes and after treated with ibuprofen or compounds for 24 hours. Then, supernatants were collected and amount of cytokines and chemokines were determined (TNFα, IL-8, IL-6, CCL3 and IL-1β in M1 cultures; and TNFα, IL-8, CCL3, IL-10, CXCL9and
CCL2in M2 cultures) using the Flow Cytomix system (eBioscience) following the instructions given by manufacturer. Gallios Beckmann Coulter cytometer was used and data was analyzed by FlowCytomixPro software, version 3.0.
CCL2in M2 cultures) using the Flow Cytomix system (eBioscience) following the instructions given by manufacturer. Gallios Beckmann Coulter cytometer was used and data was analyzed by FlowCytomixPro software, version 3.0.
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