Msd discovery workbench software

Manufactured by Mesoscale

Sourced in United States

The MSD Discovery Workbench software is a data analysis and visualization tool for researchers working with Mesoscale Discovery (MSD) technology. It provides a platform for managing, analyzing, and presenting data generated from MSD-based experiments.

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13 protocols using msd discovery workbench software

Cytokine Profiling in Mouse Neuroinflammation

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Plasma protein levels of IL-1β, IL-2, IL-6, IL-10, TNF-α, and IFN-γ were measured using a V-Plex Meso Scale Discovery (MSD) Multiplex spot assay Mouse Neuroinflammation 1 panel. All samples were diluted at a ratio of 1:4 with diluent 41—provided in the MSD kit. Samples and Standards were run in triplicates according to manufacturer instructions and analyzed with MSD Discovery Workbench software (Meso Scale Discovery, Gaithersburg, MD).

Golshiri K., Ataabadi E.A., Jüttner A.A., Snyder G.L., Davis R.E., Lin A., Zhang L., de Vries R., Garrelds I.M., Leijten F.P., Danser A.H, & Roks A.J. (2022). The Effects of Acute and Chronic Selective Phosphodiesterase 1 Inhibition on Smooth Muscle Cell-Associated Aging Features. Frontiers in Pharmacology, 12, 818355.

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Multiplex Assay for Mouse Neuroinflammation

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For mouse plasma, protein levels of IL-1β, IL-2, IL-4, IL-6, IL-10, TNF-α, and IFN-γ were measured using a V-Plex Meso Scale Discovery (MSD) multiplex spot assay mouse neuroinflammation 1 panel. All samples were diluted at a ratio of 1 : 4 with diluent 41—provided in the MSD kit. Samples and standards were run in duplicate or triplicate according to manufacturer's instructions and analyzed with MSD Discovery Workbench software (Meso Scale Discovery, Gaithersburg, MD) at ITCI.

Ataei Ataabadi E., Golshiri K., van der Linden J., de Boer M., Duncker D.J., Jüttner A., de Vries R., Van Veghel R., van der Pluijm I., Dutheil S., Chalgeri S., Zhang L., Lin A., Davis R.E., Snyder G.L., Danser A.H, & Roks A.J. (2021). Vascular Ageing Features Caused by Selective DNA Damage in Smooth Muscle Cell. Oxidative Medicine and Cellular Longevity, 2021, 2308317.

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Quantification of Inflammatory Markers

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Plasma concentrations of TNF were measured using V-PLEX Human Proinflammatory Panel 1 and plasma concentrations of TNFR1 and TNFR2 were measured using Human TNFR-I ultra-sensitive kit and Human TNFR-II ultra-sensitive kit. Interleukin (IL)-6 and vascular endothelial growth factor (VEGF-A) were measured using the V-Plex Human IL-6 kit and V-Plex Human VEGF kit (all from Mesoscale Discovery, Rockville, MD, USA). Serum concentrations of CCL2 were measured using a V-PLEX human MCP-1 kit (Mesoscale Discovery). Samples were diluted in Diluent-41 and analyzed in duplicate according to the manufacturer’s instructions. Analysis was performed on a SECTOR Imager 6000 plate reader and MSD Discovery Workbench software was used for analysis (Mesoscale Discovery). Sample replicates with coefficient of variation (CV) values >25% in individual analyses were excluded. None of the measurements were below the lower level of detection.

Hansen R.B., Laursen C.C., Nawaz N., Madsen J.S., Nielsen H.H., Kruuse C., Møller A., Degn M, & Lambertsen K.L. (2021). Leukocyte TNFR1 and TNFR2 Expression Contributes to the Peripheral Immune Response in Cases with Ischemic Stroke. Cells, 10(4), 861.

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Cytokine Profiling of Macrophages

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Supernatants were removed from maturing macrophages at day three and six post-plating. ELISA assays were performed for both IL-10 and TNF on these supernatants (Biolegend ELISA Max kits) according to manufacturer instructions. Other culture supernatants were obtained from fully differentiated macrophages after infection with Mtb, treatment with Mtb lysate, or untreated controls, followed by washout and incubation for 24h in complete media. TNF ELISAs were performed on supernatants, diluted 1:20 in assay diluent, and IL-10 ELISAs were performed neat or at a 1:1 dilution. Standard curves for each cytokine were generated in duplicate. Samples were analyzed using the Synergy H1 microplate reader (BioTek, Winooski, VT). For multiplexed cytokine analysis, a total of 37 different cytokines were analyzed in batches of 6–10 analytes per assay from macrophage culture supernatants by use of both V-PLEX and U-PLEX assays (Mesoscale Discovery, Rockville, MD) per manufacturer’s instructions. Acid treatment of sample supernatants was performed for TGFβ assays. Plates were analyzed on the MESO QuickPlex SQ 120MM and visualized on MSD Discovery Workbench software (Mesoscale Discovery).

Gail D.P., Suzart V.G., Du W., Kaur Sandhu A., Jarvela J., Nantongo M., Mwebaza I., Panigrahi S., Freeman M.L., Canaday D.H., Boom W.H., Silver R.F, & Carpenter S.M. (2023). Mycobacterium tuberculosis impairs human memory CD4+ T cell recognition of M2 but not M1-like macrophages. iScience, 26(9), 107706.

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Cytokine Profiling in Murine BAL Fluid

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Levels of IL-12p70, IL-6, KC, IL-10 and TNF-α in the BAL fluid were measured using the mouse proinflammatory panel 1 kit (Mesoscale Discovery, Gaithersburg, MD, USA), according to the manufacturer’s instructions. Data were acquired on a MESO QuickPlex SQ 120 system and analyzed with the MSD Discovery Workbench software (Mesoscale Discovery).

Heulens N., Korf H., Cielen N., De Smidt E., Maes K., Gysemans C., Verbeken E., Gayan-Ramirez G., Mathieu C, & Janssens W. (2015). Vitamin D deficiency exacerbates COPD-like characteristics in the lungs of cigarette smoke-exposed mice. Respiratory Research, 16(1), 110.

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Plasma C3, Properdin, and C5 Quantification

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Plasma C3 was measured by enzyme-linked immunosorbent assay.¹⁴ (link) Plasma “free” properdin and C5 levels were measured using electrochemiluminescence immunoassays (Meso Scale Discovery [MSD], Meso Scale Diagnostics, Rockville, MD) with anti-properdin or a custom anti-murine C5 coated on high-binding MSD plates. Bound “free” properdin and C5 were detected with anti-properdin (from W. Song) or biotinylated monoclonal anti-C5 (Alexion) combined with streptavidin-SULFO (MSD, Meso Scale Diagnostics). The chemiluminescent signal was acquired using a SECTOR S 6000 imager (MSD, Meso Scale Diagnostics) and was analyzed with MSD Discovery Workbench software (Meso Scale Diagnostics, version 4.0.12.1). Recombinant murine properdin and C5 were used as standards.

Gilmore A.C., Zhang Y., Cook H.T., Lavin D.P., Katti S., Wang Y., Johnson K.K., Kim S, & Pickering M.C. (2021). Complement activity is regulated in C3 glomerulopathy by IgG–factor H fusion proteins with and without properdin targeting domains. Kidney International, 99(2), 396-404.

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Brain Cytokine Quantification using V-PLEX

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Sample preparation for V-PLEX Proinflammatory Panel 1 Mouse kit (Meso Scale Diagnostics, Rockville, MD) was described previously [32 (link)]. Brains were collected and snap-frozen in isopentane then stored at −20 °C until time of homogenization. Brains were homogenized in RIPA buffer (Thermo Fisher Scientific, Waltham, MA) with Halt Protease and Phosphatase Inhibitor Cocktail (Thermo Fisher Scientific, Waltham, MA). Samples were centrifuged at 13,300 RPM for 15 min at 4 °C. Supernatant was collected and stored at −20 °C. Pierce BCA Protein Assay was performed to determine total protein concentration. Samples were diluted and loaded on the MSD plate at the same total protein concentrations. Manufacturer’s protocol for the V-PLEX Proinflammatory Panel 1 Mouse kit was followed as described. Plates were read with Meso Quickplex SQ 120 (Meso Scale Diagnostics, Rockville, MD) and data was analyzed via MSD Discovery Workbench software (Meso Scale Diagnostics, Rockville, MD). Concentrations of cytokines are presented as pg/mL.

Blethen K.E., Sprowls S.A., Arsiwala T.A., Wolford C.P., Panchal D.M., Fladeland R.A., Glass M.J., Dykstra L.P., Kielkowski B.N., Blackburn J.R., Andrick C.J, & Lockman P.R. (2023). Effects of whole-brain radiation therapy on the blood–brain barrier in immunocompetent and immunocompromised mouse models. Radiation Oncology (London, England), 18, 22.

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Murine Serum Cytokine Profiling

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In mouse serum, protein levels of IL-1β, IL-6, IL-10, and TNF-α were measured using a V-Plex Meso Scale Discovery (MSD) Multiplex spot assay Mouse Neuroinflammation 1 panel (Meso Scale Diagnostics). All samples were run in duplicates or triplicates according to manufacturer instructions and analyzed with MSD Discovery Workbench software (Meso Scale Diagnostics).

Dutheil S., Watson L.S., Davis R.E, & Snyder G.L. (2023). Lumateperone Normalizes Pathological Levels of Acute Inflammation through Important Pathways Known to Be Involved in Mood Regulation. The Journal of Neuroscience, 43(5), 863-877.

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Quantitative Phosphorylation Signaling in SCI

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Quantitative determination of nuclear and cytoplasmic phosphorylated (p)-NF-κB (Ser536) was investigated in naïve mice and in mice with 6 h survival after SCI using the phospho-NF-κB (Ser536) whole cell lysate kit according to manufacturer’s instructions. The phospho-STAT signaling cascade (phospho-STAT3, phospho-STAT4, phospho-STAT5a/b) and the Akt signaling cascade (phospho-Akt (Ser473), phospho-p70S6K (Thr421/Ser424), and phospho-GSK-3β (Ser9)), the ERK-STAT signaling cascade (phospho-MEK1/2 (Ser217/221), phospho-ERK-1/2 (Thr/Tyr: 202/204; 185/187), and phospho-STAT3 (Tyr705)) were investigated in naïve mice and in mice with 6 h survival after SCI according to manufacturer’s instructions. All kits were from Mesoscale. Samples were diluted prior to measurements according to manufacturer’s instructions. Samples were measured in duplicates and data was analyzed using MSD Discovery Workbench software (Mesoscale Discovery, Rockville, MD, USA).

Ellman D.G., Lund M.C., Nissen M., Nielsen P.S., Sørensen C., Lester E.B., Thougaard E., Jørgensen L.H., Nedospasov S.A., Andersen D.C., Stubbe J., Brambilla R., Degn M, & Lambertsen K.L. (2020). Conditional Ablation of Myeloid TNF Improves Functional Outcome and Decreases Lesion Size after Spinal Cord Injury in Mice. Cells, 9(11), 2407.

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Multiplexed Immunoassay Protocol

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All chemicals were purchased from Merck (Kenilworth, NJ, USA) and Sigma-Aldrich (St. Louis, MO, USA). MSD GOLD 96well Streptavidin SECTOR Plates, MSD Read Buffer T (4x), SULFO-TAG labeled goat anti-mouse IgG and QuickPlex SQ 120 system containing the MESO QuickPlex SQ 120 Imager and MSD DISCOVERY WORKBENCH software were acquired from Meso Scale Diagnostics, LLC (Rockville, MD, USA).

Tzara O., Amalie Simonsen S., Sode West A., Asser Karsdal M., Klingenberg Iversen H, & Henriksen K. (2022). Quantification of Tau-A in serum after brain injury: a comparison of two analytical platforms, ELISA and electrochemiluminescence immunoassay. Brain injury, 36(6).

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