PC3U or A549-conditioned media were concentrated 50× through Amicon Ultra-15 centrifugal filters (Merck/Millipore) at 3000× g for 15 min at 4 °C or used without concentration (for certain PC3U experiments). Secreted mature TGFβ1 was measured using the human TGFβ1-Duoset ELISA kit according to the manufacturer’s instructions (R&D Systems, Oxon, UK).
Human tgfβ1 duoset elisa kit
Manufactured by R&D Systems
Sourced in United Kingdom, United States
The Human TGFβ1-Duoset ELISA kit is a laboratory tool used for the quantitative measurement of human transforming growth factor beta 1 (TGFβ1) in biological samples. It is an enzyme-linked immunosorbent assay (ELISA) kit that provides the necessary reagents and protocols to perform this analysis.
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5 protocols using human tgfβ1 duoset elisa kit
1
Quantifying Secreted TGFβ1 from Cell Lines
2
Amperometric Immunosensor for TGF-β1 Detection
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The use of an amperometric immunosensor for the sensitive determination of TGF‐β1 has been previously described [26 ]. The immunosensor implies the implementation of a sandwich immunoassay onto the surface of carboxylic acid‐functionalized magnetic microparticles (Dynabeads M‐270 Immunoassay; Invitrogen) and coupled with screen‐printed carbon electrodes (DropSens‐Metrohm, Oviedo, Spain). Recombinant human TGF‐β1 standard, mouse anti‐TGF‐β1 capture antibody, and biotinylated chicken anti‐human TGF‐β1 detection antibody used for sandwich immunocomplex formation were components of the human TGF‐β1 DuoSet ELISA Kit (R&D Systems).
3
Quantification of Secreted BMP4 and TGFβ1
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U2987MG conditioned media were concentrated 50× through Amicon Ultra-15 centrifugal filters (Millipore) at 3000 × g for 15 min at 4 °C. Secreted BMP4 and TGFβ1 were measured using the human BMP4-ELISA Kit and the human TGFβ1-Duoset ELISA Kit, respectively (R&D Systems), according to the manufacturer’s instructions.
4
Quantifying Secreted TGFβ via ELISA
5
Quantitative Protein Analysis of Cell Supernatants
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Cultured cell supernatants were prepared and analyzed using quantikine human TIMP‐1 ELISA kit, human TGF‐β1 Duoset ELISA kit, human total MMP‐1 ELISA kit, and human MMP‐9 ELISA kit (R&D Systems, Minneapolis, MN, USA). ELISAs were conducted according to the manufacturer's instructions. All samples were analyzed in triplicates in each experiments.
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