Plasma IgA, IgG1, IgG2a, IgG2b and IgM levels were assessed with the use of Mouse Isotyping Panel 1 Assay kit from Mesoscale according to the company’s instructions (MSD Multi-spot Assays, Mesoscale Discovery, Rockville, Maryland). Plasma IgM antibodies targeting PC were detected with an ELISA kit from Athera Biotechnologies (Athera Biotechnologies AB, Solna, Sweden) according to the company’s instructions, with the exception of using peroxidase-conjugated anti-mouse IgM (Jackson ImmunoResearch, Novakemi, Handen, Sweden).
Msd multi spot assay
Manufactured by Mesoscale
Sourced in United States
The MSD Multi-spot Assays is a lab equipment product from Mesoscale. It is designed to perform simultaneous detection and quantification of multiple analytes in a single sample. The core function of this product is to enable multiplexed immunoassays, allowing researchers to analyze multiple target molecules in a cost-effective and efficient manner.
Automatically generated - may contain errors
Lab products found in correlation
Mouse isotyping panel 1 assay kit, by Mesoscale (2 mentions)
Peroxidase conjugated anti mouse igm, by Jackson ImmunoResearch (2 mentions)
Mouse ige elisa kit, by Fortis Life Sciences (1 mentions)
Heparin, by Merck Group (1 mentions)
Bat 12, by Physitemp (1 mentions)
Cd4 t cell isolation kit 2, by Miltenyi Biotec (1 mentions)
Anti cd28, by Thermo Fisher Scientific (1 mentions)
Anti cd3, by BD (1 mentions)
4 protocols using msd multi spot assay
1
Quantification of Murine Antibody Isotypes
2
Immunoglobulin Levels Quantification
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Plasma IgA, IgG1, IgG2a (not expressed in C57BL/6 mice), IgG2b, IgG3 and IgM levels were assessed with the use of Mouse Isotyping Panel 1 Assay kit from Mesoscale according to the company’s instructions (MSD Multi-spot Assays, Mesoscale Discovery, Maryland, USA). Plasma IgE levels were determined with the use of mouse IgE ELISA Kit from Bethyl Laboratories (Bethyl Laboratories Inc, Montgomery, Texas, USA) while IgM antibodies targeting phosphorylcholine were assessed with an ELISA kit from Athera Biotechnologies (Athera Biotechnologies AB, Solna, Sweden) according to the company’s instructions, with the exception of the use of peroxidase-conjugated anti mouse IgM (Jackson ImmunoResearch).
3
Quantifying Sepsis Biomarkers in Mice
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Serum cytokines were quantified by using the MSD Multi-Spot assay (Meso Scale Diagnostics, LLC) according to the manufacturer’s instructions. Blood sepsis biomarkers (pH, base deficit, and bicarbonate) were analyzed using the i-STAT system. For experiments in which i-STAT measurements were made, it was necessary to treat mice with intraperitoneal heparin (25 U; Sigma-Aldrich, St. Louis, MO) with simultaneous sedation by intraperitoneal ketamine (100 mg/kg) and xylazine (10 mg/kg) prior to cardiac puncture to prevent clotting in the i-STAT cartridges. Whole blood was drawn into a 25-gauge syringe and aliquoted into i-STAT cartridges. Values were read on an i-STAT portable clinical analyzer.
Temperatures were measured rectally using a digital thermometer (model BAT-12; Physitemp Instruments, Inc.).
Temperatures were measured rectally using a digital thermometer (model BAT-12; Physitemp Instruments, Inc.).
4
Isolation and Activation of Human CCR6+ T Cells
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Human blood samples from healthy donors were provided by the Scripps Research Institute, La Jolla following Institutional Review Board approved protocol. Total CD4+ T cells were isolated from the peripheral blood mononuclear cells (PBMCs) using a CD4+ T cell Isolation Kit II (Miltenyl Biotec, Auburn, CA), following the manufacturer’s instructions. CCR6+ cells were isolated from CD4+ T cells using phycoerythrin (PE) conjugated anti-CCR6 antibody followed by anti-PE microbeads. Isolated CCR6+ T cells were resuspended in culture medium and added to a 96-well plate which was pre-coated with 1 μg/ml anti-CD3 (BD Bioscience) at 1 × 105 cells per well. Titrated JNJ-54271074 was added to each well, then anti-CD28 (eBioscience) was added at final concentration of 5 μg/ml. Cells were cultured at 37 °C and 5% CO2 for 6 days. Supernatants were assayed for accumulated cytokines using MSD multi-spot assay (Meso Scale Discovery, Rockville, MD) or R&D ELISA kit, and cells were used for intracellular staining.
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