Cytokine (CXCL1, CXCL2, IL‐6, CXCL8, CCL2, and TNF‐α) quantification was performed using a customized all in one Bio‐Plex Pro Human Chemokine 6plx EXP kit (17002259, Bio‐Rad). The cytokines in the collected supernatants were measured by the Luminex‐based multiplex technique according to the manufacturer's instructions (Bio‐Rad). The concentrations were calculated with Bio‐Plex Manager 6.0 by comparison with the standard curves. The detection sensitivity ranged between 1 pg and 40 μg of protein per 1 ml.
Luminex based multiplex technique
Manufactured by Bio-Rad
Sourced in United States
The Luminex-based multiplex technique is a powerful analytical tool that enables the simultaneous detection and quantification of multiple analytes in a single sample. This method utilizes color-coded magnetic beads coated with specific capture antibodies or probes, allowing for the simultaneous measurement of various biomolecules, such as proteins, cytokines, or nucleic acids, in a high-throughput and efficient manner.
Automatically generated - may contain errors
Lab products found in correlation
Manager 6, by Bio-Rad (2 mentions)
Taqman 7500 real time pcr system, by Thermo Fisher Scientific (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions)
Taqman gene expression master mix, by Thermo Fisher Scientific (1 mentions)
4 protocols using luminex based multiplex technique
1
Multiplex Cytokine Quantification in Supernatants
2
Multiplex cytokine quantification
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Cytokine concentrations in supernatants of treated PMBC or isolated NK cells were quantified by the Luminex-based multiplex technique according to the manufacturer’s instructions (BioRad, Hercules, USA). Standard curves and concentrations were calculated with Bio-Plex Manager 6.1 software.
3
Multiplex Cytokine Quantification in Co-Culture
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To quantify the cytokines in the co-culture supernatants, they were centrifuged at 4,000× g for 5 min. Clear supernatants were transferred into new tubes and stored at −80 °C. The concentrations of interleukin 6 (IL-6), interleukin 8 (CXCL8), monocyte chemoattractant protein-1 (MCP-1/CCL2), tumor necrosis factor-α (TNF-α), growth regulated oncogene-α (Gro-α/CXCL1), and growth regulated oncogene-β (Gro-β/CXCL2) were quantified by the Luminex-based multiplex technique according to the manufacturer’s instructions (Bio-Rad, Hercules, USA). Standard curves and concentrations were calculated using the Bio-Plex Manager 6.0 with protein detection sensitivity between 1 pg/mL and 40 µg/mL. All samples were measured as triplicates.
4
Serum Cytokine Profiling in Lung Disease
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Concentrations of interferon (IFN)-γ, chemokine (C-X-C motif) ligand (CXCL10) (IP-10), interleukin (IL)-10, and IL-17 in serum were analyzed on POD 0 and POD 70 by using the Luminex-based multiplex technique (BioRad). Standard curves and concentrations were calculated with the use of Bio-Plex Manager 6.0. To account for interindividual variation, the baseline value at POD 0 was normalized 100% in all groups. For measurement of cytokine expression at mRNA level in lung tissue, RNA was isolated by phenol-chloroform extraction and transcribed to cDNA using the high-capacity cDNA reverse transcription kit (Applied Biosystems, Foster City, CA). Synthesized cDNA was quantified by real-time PCR (TaqMan 7500 real-time PCR system; Applied Biosystems) applying TaqMan Gene Expression Master Mix (Applied Biosystems) and TaqMan Gene Expression Assays (Applied Biosystems) specific for the cytokines and three endogenous controls: polymerase (RNA) II (Polr2A), betaglucuronidase (GUSB), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Cycle threshold values were normalized to the mean expression of the references.
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