Cytokines were preliminarily measured in cell culture supernatants (400 μL) of two samples, control and treated with the oxysterols mixture, 20 μM in ethanol, by a semi‐quantitative method to simultaneously detect the relative levels of 36 different cytokines and chemokines using the Human Cytokine Array Panel A (R&D systems, Abingdon, UK) following the manufacturer instructions. The membranes were developed using ImageQuant LAS 4000 mini. Signal intensities of each membrane array were analyzed using the ImageQuant software (Molecular Dynamics, Amersham Pharmacia Biotech). Three cytokines were selected for quantitative analysis by ELISA on the basis of the initial screening process described earlier. Levels of human cytokines IL‐1β, MIF, and RANTES were quantified using appropriate kits (“Human IL‐1β/IL‐1F2 DuoSet,” “Human MIF DuoSet,” and “Human CCL5/RANTES DuoSet” R&D systems, Abingdon, UK), following the manufacturer's instructions.
Human mif duoset
Manufactured by R&D Systems
Sourced in United Kingdom
The Human MIF DuoSet is a sandwich enzyme-linked immunosorbent assay (ELISA) kit for the quantitative measurement of human macrophage migration inhibitory factor (MIF) in cell culture supernatants, serum, and plasma. The kit includes a pre-coated capture antibody, a detection antibody, and a calibrator for quantification.
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Lab products found in correlation
Human cytokine array panel a, by R&D Systems (1 mentions)
Imagequant software, by GE Healthcare (1 mentions)
Human il 1β il 1f2 duoset, by R&D Systems (1 mentions)
Human ccl5 rantes duoset, by R&D Systems (1 mentions)
Elisa kit, by Cusabio (1 mentions)
Imark microplate absorbance reader, by Bio-Rad (1 mentions)
3 protocols using human mif duoset
1
Cytokine Profiling of Oxysterol-Treated Cells
2
Quantifying GDF-15 and MIF in MSC Secretome
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According to the manufacturer’s instructions, the concentration of GDF-15 and MIF in the CdM from iPSC-MSCs and BM-MSCs was measured using a human GDF-15 Elisa kit (Human GDF-15 DuoSet, DY957, R&D systems) and human MIF Elisa kit (Human MIF DuoSet, DY289, R&D systems) respectively.
3
Quantification of Serum MIF Levels
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Human MIF serum levels were detected by a commercially enzyme-linked immunosorbent assay (ELISA) (R&D SYSTEMS Human MIF DuoSet) kit according the protocol provided by the . manufacturer with a lower limit of 31.3 pg/mL. Serum levels of MIF in sham-operated, HS and ISO-1 treated rats from the acute model of HS were quantified using a commercially available ELISA kit (Cusabio Biotech, Wuhan, China) with a lower limit of 62.5 pg/mL by following the manufacturer/product specific protocol. Detection occurred at 450 nm and 540 nm using iMark® microplate absorbance reader (BioRad).
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