Quantification of cytokines present in the ascites fluid from mice bearing ID8-VEGF tumors was performed using the Meso Scale Discovery Multi-Spot Assay System (Meso Scale Diagnostics, LLC). V-PLEX assays were performed using Cytokine Panel 1, Proinflammatory Panel 1, and Th17 Panel 1 mouse kits, as well as a TGF-B kit, per manufacturer’s instructions. Briefly, MULTI-SPOT assay plates pre-coated with capture antibodies were washed and incubated with diluted ascites fluid samples and/or calibrators for 2 h at room temperature. Next, plates were washed and bound analytes measured using detection antibodies conjugated with electrochemiluminescent labels (MSD SULFO-TAG). Following an additional 2-h incubation at room temperature, plates were washed, incubated with electrochemiluminescence buffer, and analyzed on a MESO QuickPlex SQ 120 instrument. Data was processed using the MSD Discovery Workbench Version 4.0.
Meso scale discovery multi spot assay system
Manufactured by Mesoscale
The Meso Scale Discovery Multi-Spot Assay System is a laboratory instrument designed for multiplex immunoassay analysis. It utilizes electrochemiluminescence technology to detect and quantify multiple analytes simultaneously from a single sample. The system provides a platform for high-throughput, sensitive, and reproducible measurements of various biomolecules, including proteins, peptides, and small molecules.
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3 protocols using meso scale discovery multi spot assay system
1
Quantification of Cytokines in Ascites Fluid
2
Cytokine Profiling in Sauna Dehydration
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Plasma IL-6, IL-8, IL-10, IFN-ɣ, TNF-α concentrations, at the control visit and after sauna dehydration, were measured using a V-Plex proinflammatory panel 1, Meso Scale Discovery multi-spot assay system (MSD, Meso Scale Diagnostics, Rockville, MD) following the manufacturer’s instructions. The lower limit of quantification for this assay was specified to be of 0.063 pg/mL for IL-6, 0.591 pg/mL for IL-8, 0.298 pg/mL for IL-10, 1.76 pg/mL for IFN-ɣ and 0.690 pg/mL for TNF-α. For data analysis, values below the quantification limit were set to zero.
3
Quantification of Ovarian Cancer Cell Cytokines
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Quantification of cytokines present in the supernatant from ovarian cancer cells treated in-vitro with olaparib, Hu-5F9 or both was performed using the Meso Scale Discovery Multi-Spot Assay System (Meso Scale Diagnostics, LLC). V-PLEX assays were performed per manufacturer’s instructions. Briefly, MULTI-SPOT assay plates pre-coated with capture antibodies were washed and incubated with diluted supernatant fluid samples and/or calibrators for two hours at room temperature. Next, plates were washed and bound analytes measured using detection antibodies conjugated with electrochemiluminescent labels (MSD SULFO-TAG). Following an additional two hours incubation at room temperature, plates were washed, incubated with electrochemiluminescence buffer, and analyzed on a MESO QuickPlex SQ 120 instrument. Data was processed using MSD Discovery Workbench Version 4.0.
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