The concentrations of human IFN-β (VeriKine™ Human IFN β ELISA; PBL Assay Science, Piscataway, NJ), IL-6 (Human IL-6 ELISA MAX™ Deluxe, Biolegend, San Diego, CA), IL-7 (Human IL-7 Quantikine HS ELISA Kit; R&D systems, Minneapolis, MN), and IL-15 (Human IL-15 Quantikine ELISA Kit; R&D systems) in conditioned media from human MSCs were measured in accordance with the manufacturer's protocols. All samples were examined in triplicate for each experiment.
Human il 7 quantikine hs elisa kit
Manufactured by R&D Systems
Sourced in United States
The Human IL-7 Quantikine HS ELISA kit is a solid-phase enzyme-linked immunosorbent assay (ELISA) designed to measure human interleukin-7 (IL-7) levels in cell culture supernates, serum, and plasma.
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Lab products found in correlation
Human il 6 elisa max deluxe, by BioLegend (1 mentions)
Human il 15 quantikine elisa kit, by R&D Systems (1 mentions)
Elisa max deluxe set, by BioLegend (1 mentions)
Automacs pro separator, by Miltenyi Biotec (1 mentions)
Human il 6 elisa ready set go, by Thermo Fisher Scientific (1 mentions)
Infinite m200 elisa reader, by Tecan (1 mentions)
Magellan version 7, by Tecan (1 mentions)
Infinite m200, by Tecan (1 mentions)
7 protocols using human il 7 quantikine hs elisa kit
1
Quantifying Cytokine Secretion in Human MSCs
2
Quantification of IL-7 in Cell Culture
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The concentrations of IL-7 in culture media were measured by ELISA. The culture media were collected after centrifugation for cell pelleting, and the levels of IL-7 were quantified by using the Human IL-7 Quantikine HS ELISA Kit (HS750; R&D Systems) according to the manufacturer’s instructions.
3
Cytokine Profiling in Cell Therapy
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Patients’ serum was collected on day 0, before cell infusion. IL-7, IL-15 and IL-21 serum levels were determined by ELISA (IL-7, Human IL-7 Quantikine HS ELISA kit, R&D Systems, Minneapolis, Minnesota, USA; IL-15 and IL-21, ELISA MAX Deluxe Set, BioLegend, San Diego, California, USA) according to the manufacturer’s instructions. Measurements were performed in duplicates.
4
Quantification of IL-7 in Macrophage-TE Cell Co-cultures
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Macrophages, and TE-9, -10, and -11 cells (1 × 106 cells/well) in RPMI-1640 with 10% FBS were cultured in 6-well plates for 48 h. In addition, after separation using an autoMACS Pro Separator (Miltenyi Biotec), monocultured and co-cultured TE-9, -10, and -11 cells (3 × 105 cells/well) in RPMI-1640 with 10% FBS were cultured in 6-well plates for 48 h. The cell culture supernatants were collected via a previously described method [19 (link)]. The concentrations of IL-7 in the cell culture supernatants were measured using the Human IL-7 Quantikine HS ELISA Kit (#HS750; R&D Systems) according to the manufacturer’s instructions. The optimal density of each well was determined at 490 nm using an Infinite 200 PRO microplate reader.
5
Quantification of IL-6 and IL-7 in Plasma
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IL-6 and IL-7 was determined in duplicate for diluted plasma samples using Human IL-6 ELISA Ready-SET-Go! (eBioscience) and Human IL-7 Quantikine HS ELISA kit (R&D Systems), respectively, according to manufacturer’s instructions. Samples were measured using the Infinite M200 ELISA reader (Tecan). Concentrations were calculated from the respective standard curves by applying 4-parametric logistic regression. Samples outside the detection range were set to the corresponding lower or upper range value.
6
Cytokine production and IL-7 quantification
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Cytokine production at the single-cell level was assessed after 4 hours stimulation with PMA+Ionomycin and Brefeldin A, as described [44 (link)]. Serum IL-7 levels were quantified using Human IL-7 Quantikine HS ELISA kit (R&D Systems) [42 (link)].
7
Quantification of Serum IL-7 Levels
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Serum of study participants was stored at -80 °C for concomitant IL-7 measurement at the University Childrens Hospital in Duesseldorf. Serum IL-7 concentrations were measured as described previously [16, 20] . In brief, we used a commercially available highly sensitive ELISA (Human IL-7 Quantikine HS ELISA kit, R&D Systems) according to manufacturer's instructions. Because of low IL-7 serum concentrations, we included standards on each plate to increase the accuracy of these assays. All samples were measured at least as duplicates. Optical density was determined using an ELISA reader (Infinite M200, Tecan). IL-7 concentrations were calculated from the standard curve by applying 4-parametric logistic regression (Magellan version 7.2, Tecan).
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