Cytokine levels (IL-4, IL-10, IL-13 and IL-6) in CM from YUMM1.7 or MEF cells were measured using the kit LEGENDplex murine Th Cytokine panel 12-plex, according to manufacturer’s instruction. Samples were acquired on LSRII and analysed using LEGENDplex software (BioLegend). Cholesterol and fatty acid in CM and CM without lipids were measured by using Total Cholesterol and Cholesteryl Ester Colorimetric/Fluorometric Assay Kit (Biovision) and Free Fatty Acid Quantification Colorimetric/Fluorometric Kit (Biovision) respectively. Quantification was performed following manufacturer’s instructions.
Legendplex software
Manufactured by BioLegend
Sourced in United States
The LEGENDplex software is a data analysis tool designed to facilitate the processing and interpretation of data generated from multiplex assays. The software provides users with the ability to analyze and visualize the results of their experiments.
Automatically generated - may contain errors
Lab products found in correlation
Legendplex mouse inflammation panel, by BioLegend (8 mentions)
Legendplex multi analyte flow assay kit, by BioLegend (6 mentions)
Facsverse, by BD (6 mentions)
Facsdiva software, by BD (4 mentions)
Facscanto 2, by BD (4 mentions)
Legendplex, by BioLegend (4 mentions)
Lsrfortessa 5, by BD (4 mentions)
Sa pe, by BioLegend (3 mentions)
Facsaria 3 flow cytometer, by BD (3 mentions)
Lsr 2, by BD (3 mentions)
Lsrii flow cytometer, by BD (3 mentions)
Legendplex kit, by BioLegend (3 mentions)
Protease inhibitor cocktail, by Merck Group (3 mentions)
Mouse proinflammatory chemokine panel, by BioLegend (3 mentions)
Protease inhibitor cocktail, by Roche (3 mentions)
Lsrfortessa, by BD (3 mentions)
Free fatty acid quantification colorimetric fluorometric kit, by Abcam (2 mentions)
Total cholesterol and cholesteryl ester colorimetric fluorometric assay kit, by Abcam (2 mentions)
Legendplex human inflammation panel 1, by BioLegend (2 mentions)
Legendplex human type 1 2 3 interferon panel, by BioLegend (2 mentions)
106 protocols using legendplex software
1
Quantifying Cytokines and Lipids in CM
2
Cytokine and Immunoglobulin Profiling
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Mouse serum and supernatant of cultured splenocytes were collected and stored at −80°C for detection. Cytokines in serum were measured using an LEGENDplex multianalyte flow assay kit according to the manufacturer’s protocol, and the concentration of cytokines was determined by using LEGENDplex software (BioLegend). Immunoglobulin level in cultured splenic B cell supernatant was detected by cytometric bead array (CBA) assay according to the manufacturer’s protocol and read by flow cytometry (Cystek, Fremont, CA, USA).
3
Multiplex Cytokine Quantification in Mice
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Cytokines were quantified using a LEGENDplex multianalyte flow assay kit (BioLegend, San Diego, CA). Serum from wild-type, αβR−/−, λR−/−, and αβR−/− λR−/− mice was incubated for 2 h at room temperature with customized premixed beads and detection antibodies specific for a panel of 12 murine cytokines (IL-23, IL-12, IFN-γ, TNF-α, KC, MCP-1, IL-1β, IP-10, IL-6, IL-33, IFN-β, and GM-CSF). Samples were then incubated for 30 min with SA-PE (BioLegend) and washed, and resulting fluorescent signals were analyzed on a flow cytometer (LSRII; BD Biosciences) according to the manufacturer’s instructions. Analyte concentrations were determined and heat maps were produced using LEGENDplex software (v7.0; BioLegend, San Diego, CA).
4
Multiplex Assay for Colon Tissue
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Appropriate colon samples were mechanically disrupted in tissue extraction reagent (Thermo Fisher Scientific, 78,510) containing protease and phosphatase inhibitors (Bimake, B14001, B15001), followed by homogenization in a low-temperature grinder. The homogenate was lysed on ice for 2 h and centrifuged at 12,000 g for 10 min to extract colon tissue homogenates (CTHs). CTHs were quantified by a BCA protein quantification kit (Yeasen, 20201ES76) according to the manufacturer's instructions. Then, 50 μg of CTH was subjected to multiplex assays, according to the modified protocols from the manufacturer (Biolegend, 740,005, 740,007, 740,134). Samples were examined on a BD FACS Canto II (Beckman Coulter, Inc., Brea CA) and analyzed with LEGENDplex software (Biolegend).
5
Inflammatory Cytokine Secretion by Macrophages
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A bead-based human inflammatory cytokine array was performed using LEGENDplex human inflammation panel 1 (BioLegend). Human primary macrophages were cultured for 24 h in the presence of S protein, with or without various doses of Roneparstat (50 to 200 μg/mL). Twenty-five microliters of the conditioned medium from various conditions was used to measure the secreted cytokines in each sample, according to the instructions from the manufacturer. The measurement was performed on a FACSCalibur flow cytometer (BD), and the data were analyzed using LEGENDplex software (BioLegend).
6
Quantification of Type 1/2/3 Interferons
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Culture supernatants were stored at −80°C and thawed prior to quantification. Concentrations of IFNα2, IFNβ, IFNγ, IFNλ1, and IFNλ2/3 were measured using LEGENDplex Human Type 1/2/3 Interferon Panel (#740396; BioLegend) according to the manufacturer’s protocol. Data were acquired on a BD FACSVerse (BD) and analyzed with LEGENDplex Software (BioLegend). In figures, dotted lines indicate limits of detection.
7
Multiplex Cytokine Profiling of Tumor Cells
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Supernatants from tumor cell lysis experiments were collected after 16 h and stored at −20°C. A multiplex assay (LEGENDplex™ Human CD8/NK Panel (13-plex), BioLegend) was used according to the manufacturer's instructions. LEGENDplex™ Software from BioLegend was used for analysis of acquired data.
8
Multiplex Cytokine Profiling in Mice
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Serum concentrations of cytokines: IL-1α, IL-1β, IL-6, IL-10, IL-12p70, IL-17α, IL-23, IL-27, MCP-1, interferon-β (IFN-β), IFN-γ, tumor necrosis factor-α (TNF-α), and GM-CSF were measured using LEGENDplex Mouse Inflammation Panel (13-plex) immunoassay (Biolegend; San Diego, CA, USA) and BD LSRFortessa flow cytometer and analyzed using LEGENDplex software (Biolegend).
9
Cytokine Quantification by LEGENDPlex
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Cytokine concentrations of infection supernatants were measured via LEGENDPlex human inflammation panel (13-plex) kit (BioLegend) according to the manufacturer's instructions. Data were acquired with a FACSAria III flow cytometer using FACS DIVA Software (both BD Bioscience) and analyzed using LEGENDPlex software (BioLegend).
10
Multiplex Analysis of Inflammatory Cytokines
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Cytokines' (IL-1α, IL-1β, IL-6, IL-10, IL-12p70, IL-17α, IL-23, IL-27, MCP-1, IFN-β, IFN-γ, TNF-α, and GM-CSF) concentrations in sera were determined using LEGENDplex Mouse Inflammation Panel (13-plex) kit (Biolegend) and the BD LSRFortessa flow cytometer. The results were analyzed using LEGENDplex software (Biolegend).
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