The medium of the hMADS cells was collected over 24 h, from day 13 (after replacement of medium) to day 14 of differentiation, to determine the secretion of adipokines using high-sensitive ELISAs (leptin and DPP-4 from R&D Systems, Inc., Minneapolis, MN, USA; IL-6 and MCP-1 from Diaclone SAS, Besancon Cedex, France; adiponectin and PAI-1 from BioVendor–Laboratorni medicina a.s., Brno, Czech Republic). If necessary, samples were diluted with the dilution buffer provided by the manufacturer prior to the assay, which was performed in duplicates according to the manufacturer’s instructions.
Mcp 1
Manufactured by Diaclone
Sourced in United States, China
The MCP-1 is a laboratory instrument used for the detection and quantification of monocyte chemoattractant protein-1 (MCP-1), a chemokine involved in the recruitment and activation of monocytes and macrophages. The core function of the MCP-1 is to measure the concentration of this specific protein in various biological samples, such as cell culture supernatants, tissue extracts, or body fluids.
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2 protocols using mcp 1
1
Adipokine Secretion in hMADS Cells
2
Quantification of Plasma Biomarkers by ELISA
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Sandwich enzyme-linked immunosorbent assay (ELISA) was used for the estimation of various molecules. Commercially available ELISA kits were used to estimate the protein levels for ANG, VEGF, VEGFR2, OPTN, TDP-43 (Qayee Bio-Technology Co., Ltd., Shanghai, China), and MCP-1 (Diaclone SAS, Besancon, France) in plasma of participants as per the standard protocol described by manufacturer. Briefly, 50 µL of standard and diluted samples (range: 2–10 times of dilution) were added to the wells, after which HRP conjugated antibody was added. The plate was set for incubation at 37°C for 1 hour. This allowed the antigen to bind with antibody precoated in the wells of ELISA plate. After washing the plate, 50 µL of chromogen solution A and B were added in dark and the plate was incubated for 10 minutes at 37°C in dark. The stop solution was added and estimation was done using ELISA plate reader (iMARK reader, BioRad) at 450 nm. OD values were noted.
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