For TGF-β1 protein concentration in culture supernatants the Quantikine ELISA kit for Human TGF-β1 (R&D Systems®, Minneapolis, MN, USA) was used according to the instructions provided by manufacturers. The mean of the minimum detectable dose is 4.61 pg/mL. The TGF-β1 concentration was evaluated in control ASM cell culture supernatants; ASM cells and asthmatic eosinophils as well as ASM cells and healthy eosinophils cultures’ supernatants before and 24 h after bronchial allergen challenge. As the manufacturer notes, the Sample Activation Kit was used. The optical density was determined within the 30 min using a microplate reader set to 450 nm and for wavelength correction, the 540 nm wavelength was set to correct the optical imperfections in the plate. Every sample was done in two replicates. The results are shown as the median (range).
Quantikine elisa kit for human tgf β1
Manufactured by R&D Systems
Sourced in United States
The Quantikine ELISA kit for Human TGF-β1 is a solid-phase enzyme-linked immunosorbent assay (ELISA) designed to quantitatively measure human transforming growth factor beta 1 (TGF-β1) levels in cell culture supernatants, serum, and plasma.
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4 protocols using quantikine elisa kit for human tgf β1
1
Quantitative TGF-β1 Measurement in Cell Cultures
2
Quantifying TGF-β1 and TGF-β2 Secretion
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Cells (3 × 105 cells/well) were seeded in six-well plates (Corning Costar), and incubated for 30 h. Then, the cells were incubated in fresh FBS-free medium with DMSO (0.1%; control) or efatutazone (10 μmol/L) for an additional 60 h, and the culture medium was collected and centrifuged. The amount of TGF-β1 or TGF-β2 in the conditioned medium was quantified using the Quantikine ELISA Kit for human TGF-β1 or TGF-β2 (R&D Systems, Minneapolis, MN, USA), respectively. Then, acid activation was performed according to the manufacturer's instructions. The raw data were normalized to cell number.
3
Quantifying TGF-β1 in Cell Cultures
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A Quantikine ELISA kit for human TGF‐β1 (R&D Systems, Inc.) was used to quantify the TGF‐β1 content in the supernatants of CC cell cultures. To neutralize the biological activity of TGF‐β1 produced in the cell cultures, anti‐TGF‐β1, anti‐TGF‐β2, and anti‐TGF‐β3 neutralizing antibodies (anti‐TGF‐β, R&D) were added following the manufacturer's protocol.
4
Quantifying Serum TGF-β1 Levels
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Serum transforming growth factor β1 (TGF-β1) concentrations were measured using Quantikine® ELISA Kit for human TGF-β1 (R&D Systems, USA), according to the manufacturer’s protocol. Cytokine concentration was calculated against a standard curve.
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