TNF‐α and MCP‐1 levels in liver tissue were assessed in a Luminex 100 system (Luminex Corp., Austin, TX, USA) using a custom‐made MILLIPLEX MAP Mouse High Sensitivity T Cell Magnetic Bead Panel (Merck Millipore, Billerica, MA, USA). Briefly, 25 μL of diluent and 25 μL of liver protein extract were added to each well before the addition of premixed microbeads (25 μL). The plate was incubated overnight at 4°C with shaking, then washed and reincubated with 25 μL of detection antibody for 1 h. The plate was washed again and incubated with 25 μL of streptavidin–phycoerythrin for 30 min. The plate was finally washed twice, and the beads were resuspended with 150 μL of sheath fluid and analysed in the Luminex 100 system. Readouts were detected as mean fluorescence intensity by the instrument, and values were subsequently converted to pg·mL−1 by extrapolation from a set of standards that were run simultaneously in the assay.
Milliplex map mouse high sensitivity t cell magnetic bead panel
Manufactured by Merck Group
Sourced in United States
The MILLIPLEX MAP Mouse High Sensitivity T Cell Magnetic Bead Panel is a multiplex assay kit designed for the detection and quantification of multiple mouse T cell-related analytes in a single sample. The kit utilizes magnetic beads coated with specific capture antibodies to measure the concentrations of the target analytes.
Automatically generated - may contain errors
Lab products found in correlation
Luminex 200 platform, by Merck Group (1 mentions)
Ionomycin, by Thermo Fisher Scientific (1 mentions)
Human t activator cd3 cd28, by Thermo Fisher Scientific (1 mentions)
Anti f4 80, by Cell Signaling Technology (1 mentions)
Nu7441, by Selleck Chemicals (1 mentions)
Gapdh antibody, by Thermo Fisher Scientific (1 mentions)
6 protocols using milliplex map mouse high sensitivity t cell magnetic bead panel
1
Quantifying Inflammatory Markers in Liver Tissue
2
Liver Inflammation Biomarker Assay
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Serum concentrations of glucose, total cholesterol, TAG, AST, and ALT were determined by standard laboratory procedures. Levels of IL-1β in liver tissue were assessed in a Luminex 100 System using a custom-made MILLIPLEX MAP Mouse High Sensitivity T Cell Magnetic Bead Panel (Merck Millipore). IL-1β levels in cell supernatants were assessed by enzyme-linked immunosorbent assay (ELISA) (Beckton Dickinson) (SI Appendix ).
3
Quantifying Cytokine Profiles in Mice and Humans
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Cytokines in mice sera, DCs, or mouse DC supernatants were quantified using multiparametric Luminex kits. In brief, interferon gamma (IFN-γ), IL-2, IL-4, IL-6, IL-10, IL-12 (p70), IL-17A, KC/CXCL1, MIP-2, and TNF-α levels in mice serum samples were quantified using the Luminex 200 platform with a magnetic system (Milliplex MAP Mouse High Sensitivity T Cell Magnetic Bead Panel, EMD Millipore Corporation, Billerica, MA, USA) following the manufacturer's instructions. Cytokine concentrations are expressed as the average of three replicates in pg/mL ± SD. Similarly, cytokines in the human sera of COVID-19 patients, and vaccinated or control volunteers were quantified using the multiparametric Luminex kit (Milliplex human 1xl HSTCMAG-28SK including the following cytokines: IFN-γ, IL-10, IL-17A, IL-2, IL-4, IL-6, IL-8, TNF-α, EMD Millipore Corporation) following the manufacturer´s instructions.
4
Multiplex Cytokine Profiling in Mouse Samples
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Multiplex cytokine assay (EMD Millipore’s MILLIPLEX® MAP Mouse High Sensitivity T Cell Magnetic Bead Panel, Millipore Corp., Billerica, MA) was used for the simultaneous quantification of any of the following 18 mouse cytokines: granulocyte- macrophage colony stimulating factor (GM-CSF), interferon γ (IFN-γ), interleukin 1α (IL-1α), interleukin 1β (IL-1β), interleukin 2 (IL-2), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 7 (IL-7), interleukin 10 (IL-10), interleukin 12 (IL-12 (p70)), interleukin 13 (IL-13), interleukin 17A (IL-17A), keratinocyte-derived chemokine (KC), LPS-induced CXC Chemokine (LIX), monocyte chemotactic protein 1 (MCP-1), macrophage inflammatory protein 2-α (MIP-2) and TNF-α in mouse serum and lung tissue supernatant samples according to the kit-specific protocols. More details can be found in previous studies [16] (link), [17] (link).
5
Multiplex Cytokine Profiling in Serum
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The level of cytokines in serum was evaluated using a cancer biomarker panel of Milliplex® MAP Mouse High Sensitivity T Cell Magnetic Bead Panel (Millipore) according to the manufacturer's instructions. Peripheral blood was collected from the retrobulbar venous plexus and centrifuged at 800 g at 4°C for 15 min. The resulting supernatant was collected and 100 μl of each sample was added to a 96‐well plate in triplicate. Next, EMD Millipore beads were coated with the following cytokines: IL‐1β, IL‐2, IL‐13, IL‐10, IL‐4, IL‐6, granulocyte‐macrophage colony‐stimulating factor (GM‐CSF), TNF‐α, and IFN‐γ. The beads were then added to the serum samples and incubated with agitation at 4°C overnight. After incubation, the wells were supplemented with sheath fluid and analyzed using a multiplexing instrument. The fluorescence intensity of the samples was measured by FLEXMAP 3D® System (Luminex) and the density of the cytokines was calculated with a 5‐parameters logistic fitting curve. The production of serum cytokines was measured using Luminex® 200™ Instrument System (Invitrogen) and calculated using a software package of xPONENT (Luminex Corporation).
6
Flow Cytometry and Immunoblotting for T-cell Analysis
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