Interleukin-1β in placenta and plasma was measured by enzymelinked immunosorbent assay (ELISA) (Abcam, Cambridge, MA, USA). Following the manufacturer’s protocol, the plates were read using the CLARIOstar system (BMG Labtech, Cary, NC, USA), and final data were normalized by protein concentration.
Clariostar system
Manufactured by BMG Labtech
Sourced in United States, Germany
The CLARIOstar is a multimode microplate reader from BMG LABTECH. It is designed to measure a wide range of assays and samples in microplates. The CLARIOstar utilizes various detection modes, including absorbance, fluorescence, and luminescence, to provide comprehensive analytical capabilities.
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7 protocols using clariostar system
1
Placental Interleukin-1β Measurement
2
Screening of Metal Ions Using CLARIOstar
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The screening of metals ions was performed by using a CLARIOstar system, BMG Labtech (220–900 nm) in water from the Proteomass Scientific Society–BIOSCOPE Facility Lab. The studies were carried out by preparing aqueous solutions of metals with concentrations between 1.10−3 and 1.10−4
m . Concentrations of NPs were normalized to have an absorbance equal to 0.2. By using the 96‐well‐plate CLARIOstar system, the NPs were added to wells with the required amount of each metal.
3
Quantifying BAFF and IgG1 in Plasma
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The levels of soluble B cell-activating factor (BAFF) and IgG1 in patient plasma samples were measured by commercially available enzyme-linked immunosorbent assay (ELISA) (DBLYS0B R&D Systems, Minneapolis, USA and 88-50560-22, Invitrogen, CA, USA, respectively). The plates were read using the CLARIO star system following the manufacturer's recommended procedures (BMG Labtech, Cary, NC, USA).
4
MTT Cell Proliferation Assay
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A total of 5000 Adh and Susp cells was seeded on a 96 well plate at least 16 h prior to adding the MTT reagent.
The assay was performed with the CellTiter 96® Non-Radioactive Cell Proliferation kit (Promega Corporation, Wisconsin, USA) by following the protocol of the supplier. Afterwards, plates were analyzed by measuring the absorbance at 490 and 570 nm using the ClarioStar system (BMG LABTECH GmbH, Ortenberg, Germany).
The assay was performed with the CellTiter 96® Non-Radioactive Cell Proliferation kit (Promega Corporation, Wisconsin, USA) by following the protocol of the supplier. Afterwards, plates were analyzed by measuring the absorbance at 490 and 570 nm using the ClarioStar system (BMG LABTECH GmbH, Ortenberg, Germany).
5
Melanin Quantification in B16-F10 Cells
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Melanin content in the Susp and Adh population of B16-F10 cells were quantified as per the protocol described in [48] . For this purpose, 3D soft agarose culture assays were set up using phenol red-free RPMI (ThermoFischer Scientific, 11,835,030) complete cell culture medium. On the final day of the soft agarose colony formation assay, the culture supernatant was assessed for solubilized pigment levels at 490 nm using the ClarioStar system (BMG LABTECH GmbH, Ortenberg, Germany). The absorbance was averaged from three wells, and each experiment was performed in pentaplicates.
6
Cell Proliferation ELISA Assay
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A total of 10,000 Adh and Susp cells were seeded on a 96 well plate. The assay was carried out with the Cell proliferation ELISA, BrdU kit (Merck KgaA, Darmstadt, Germany), in accordance with the manufacturer's protocol. Plates were analyzed by measuring the absorbance at 450 and 690 nm using the ClarioStar system (BMG LABTECH GmbH, Ortenberg, Germany).
7
Placental Fibrinogen and Fibrin Analysis
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Fibrinogen levels can be used as an indirect measure for the consumption of clotting factors following thrombus formation. Fibrinogen and fibrin in placenta were measured using an enzyme-linked immunosorbent assay (ELISA) kit purchased commercially (Abcam, Cambridge, MA, USA). Following the manufacturer’s protocol, the plates were read using the CLARIOstar system (BMG Labtech, Cary, NC, USA), and final data were normalized by protein concentration. Histochemical staining (rapid phosphotungstic acid hematoxylin, PTAH, Polyscience Inc, Warrington, PA, US) was applied to the placenta for fibrin staining, according to the manufacturer's protocol.
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