Supernatant from cultured PBMCs with epcoritamab or B12 control was collected after 7 days of culture and analyzed with the Milliplex MAP Human High Sensitivity T-Cell Magnetic Bead Panel (Millipore Corporation) following the manufacturer’s protocol (supplemental Methods ).
Milliplex map human high sensitivity t cell magnetic bead panel
Manufactured by Merck Group
Sourced in United States
The MILLIPLEX® MAP Human High Sensitivity T Cell Magnetic Bead Panel is a multiplex assay designed to quantify the concentration of various cytokines and chemokines related to T-cell function. The panel utilizes magnetic beads coated with specific capture antibodies to measure the levels of these analytes in biological samples with high sensitivity.
Automatically generated - may contain errors
8 protocols using milliplex map human high sensitivity t cell magnetic bead panel
1
Cytokine Analysis of Activated PBMCs
2
Cardiac Injury Biomarkers Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Markers of cardiac injury, inflammatory response and oxidative stress were measured in blood samples collected preoperatively (prior to CPB) and postoperatively at 1, 6, 12, 24, 48, and 72 h. Whole blood samples were coagulated with EDTA and centrifuged for 10 min at 1,000 × g within 30 min of collection. Plasma was removed and assayed immediately in accordance with kit manufacturer instructions or aliquoted for storage at −20°C until use. Details are shown elsewhere (24 (link), 25 (link)). In brief, IL-6, IL-8, IL-10, TNFα and Myeloperoxidase (MPO) were measured using the Milliplex MAP Human High Sensitivity T Cell Magnetic Bead Panel (Millipore, Billerica, USA). Troponin I was measured as per standard clinical practice.
3
Cytokine Profiling for T Cell Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pro-inflammatory cytokines IL-1β, TNF-α, and IL-6 were detected by multiple analyte profiling technologies (MILLIPLEX ® MAP Human High Sensitivity T Cell Magnetic Bead Panel, EMD Millipore Corporation, Missouri, USA). For IL-1β and TNF-α, the inter-and intra-assay coefficients of variation were ≤15% and ≤5%, respectively, with sensitivity of 0.14 pg/mL for IL-1β and of 0.16 pg/mL for TNF-α. The inter-and intra-assay coefficients of variation for IL-6 were ≤20% and ≤5%, respectively, with a sensitivity of 0.11 pg/mL.
4
Analyzing Cell Secretome and Cytokine Profiles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To obtain the cell secretome, cells were seeded in a 48-well plate. At 80% confluence, they were washed 3 times and the medium was replaced with serum-free DMEM. After 48 h, the medium was collected and centrifuged at 1000× g for 10 min. The supernatant was stored at -80°C until use.
To analyze the levels of cytokines present in the secretome released by the cells, the supernatants were tested using MILLIPLEX ® MAP Human High Sensitivity T Cell Magnetic Bead Panel (EMD Millipore Corp). The cytokines and chemokines analyzed were ITAC(CXCL11), GM-CSF, Fractalkine, IFNγ, IL-10, MIP-3α, IL-12, IL-13, IL-17A, IL-1β, IL-2, IL-21, IL-4, IL-23, IL-5, IL-6, IL-7, IL-8, MIP-1α, MIP-1β, and TNF-α. The experiments were carried out following the manufacturer's instructions and the levels of cytokines were detected by MAGPIX ® technology (Millipore, MA, USA), with all samples analyzed in triplicate.
To analyze the levels of cytokines present in the secretome released by the cells, the supernatants were tested using MILLIPLEX ® MAP Human High Sensitivity T Cell Magnetic Bead Panel (EMD Millipore Corp). The cytokines and chemokines analyzed were ITAC(CXCL11), GM-CSF, Fractalkine, IFNγ, IL-10, MIP-3α, IL-12, IL-13, IL-17A, IL-1β, IL-2, IL-21, IL-4, IL-23, IL-5, IL-6, IL-7, IL-8, MIP-1α, MIP-1β, and TNF-α. The experiments were carried out following the manufacturer's instructions and the levels of cytokines were detected by MAGPIX ® technology (Millipore, MA, USA), with all samples analyzed in triplicate.
5
Inflammatory Biomarkers in Plasma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After an overnight fast (at least 12 h), blood samples were drawn from the antecubital vein. Blood samples in tubes containing EDTA were spun immediately at 1000g for 10 min. Plasma was isolated and stored at -80°C until analysis in the Center of Biomedical Research (Granada, Spain). Five key inflammatory biomarkers analysed in plasma were included in this study: white blood cell (WBC, 10 3 /µL) count, interleukin-6 (IL-6, pg/mL), interleukin-1β (IL-1β, pg/mL), tumor necrosis factor-α (TNF-α, pg/mL), and C-reactive protein (CRP, mg/L). WBC count was analysed with automated blood cell counters. IL-6, IL-1β and TNF-α were quantified by multiple analyte profiling technology (MILLIPLEX ® MAP Human High Sensitivity T Cell Magnetic Bead Panel, EMD Millipore Corporation, Missouri, U.S.A.) using a kit plex (HCYIL6-MAG Anti-Human IL-6 Beads set, HCYIL1B-MAG Anti-Human IL-1β
Bead, and HCYTNFA-MAG Anti-Human TNFα Beads set). The intra-and inter-assay precision coefficients of variation (CVs) for IL-6 were 5% and 20%, respectively, and sensitivity was 0.11 pg/mL. For both, IL-1β and TNF-α the intra-and inter-assay precision CVs were 5% and 15%, respectively, with a sensitivity of 0.14 pg/mL for IL-1β, and of 0.16 pg/mL for TNF-α. CRP was determined by turbidimetry.
Bead, and HCYTNFA-MAG Anti-Human TNFα Beads set). The intra-and inter-assay precision coefficients of variation (CVs) for IL-6 were 5% and 20%, respectively, and sensitivity was 0.11 pg/mL. For both, IL-1β and TNF-α the intra-and inter-assay precision CVs were 5% and 15%, respectively, with a sensitivity of 0.14 pg/mL for IL-1β, and of 0.16 pg/mL for TNF-α. CRP was determined by turbidimetry.
6
Luminex-based Measurement of IFN-γ and IL-12
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum Interferon-gamma (IFN-γ) and interleukin-12p70 (IL-12) concentrations were measured using the Milliplex MAP human high sensitivity T cell magnetic bead panel (Merck Millipore) on a Luminex 200 analyzer (Luminex Corporation, Austin, Texas) according to the manufacturer’s recommendations. Data analysis was performed using the BioPlex manager software v 6.1 (Bio-Rad).
7
Multiplex Immunoassay for IL-4 and IL-21 Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Determinations of IL-4 and IL-21 were performed using a multiplex bead-based immunoassay (MILLIPLEX® MAP human high-sensitivity T cell magnetic bead panel; Merck EMD Millipore, Billerica, MA) following the manufacturer’s instructions. The assay was performed on the Luminex® MAGPIX® System detection instrument operated with xPONENT Software V4.2 (both from Luminex Corp., Austin, TX). The results were analyzed using the Belysa™ Analysis software V1.2.0 (Merck KGaA, Darmstadt, Germany).
8
Multiplex cytokine profiling of samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plasma cytokines concentrations were determined using enzyme linked immunosorbent assays. Interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin 1 beta (IL-1β), tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), macrophage inflammatory proteins 1 alpha (MIP-1α) and 1 beta (MIP-1β) were analyzed using a multiplex bead based immunoassay (MILLIPLEX® MAP Human High Sensitivity T Cell Magnetic Bead Panel, Merck KgaA, Darmstadt, DE, USA). Interferon gamma induced protein 10 (IP-10) (Human CXCL10 ELISA kit, Abcam, Cambridge, UK) and soluble receptor of interleukin 2 (sCD25) were analyzed using the Human Quantikine Immunoassay (R&D Systems, Minneapolis, MN, USA). All followed the manufacturer’s instructions.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!