RAW-Blue NF-κB cells (Invivogen) were passaged and plated in a 96 well plate at 100k cells/well in 180 μL DMEM containing 10% HIFBS. Cells were incubated at 37°C and 5% CO2 for 24 h. 100 ul of cells were incubated with varying ratios of MPs at 37°C and 5% CO2 for 18 h. After 18 h, 20 μL of the cell supernatant was placed in 180 μL freshly prepared QuantiBlue (Invivogen) solution and incubated at 37°C/5% CO2 for up to 2 h. The plate was analyzed every hour using a Multiskan FC plate reader (Thermo Scientific) and absorbance was measured at 620 nm BCA Assay- This was performed according to manufacturer’s instruction (Thermo Fischer) with some modifications. 100 million beads were incubated with BCA solution and reacted for 30 mins at 60°C then analyzed every hour using a Multiskan FC plate reader (Thermo Scientific) and absorbance was measured at 562 nm and compared to a standard curve of modified MPLA or Pam2 after subtracting a background of maleimide modified MP.
Multiskan fc plate reader
Manufactured by Thermo Fisher Scientific
Sourced in United States, France, United Kingdom, China
The Multiskan FC plate reader is a versatile and reliable instrument designed for performing a wide range of absorbance-based assays in microplates. It features a robust and compact design, making it suitable for use in various laboratory settings. The Multiskan FC offers accurate and reproducible measurements across a broad wavelength range, enabling researchers to conduct a variety of spectrophotometric analyses with confidence.
Automatically generated - may contain errors
Lab products found in correlation
Quanti blue, by InvivoGen (9 mentions)
Raw blue nf κb cells, by InvivoGen (4 mentions)
Celltiter 96 aqueous one solution reagent, by Promega (2 mentions)
Non fat milk, by neoFroxx (2 mentions)
Taxol, by Merck Group (2 mentions)
Prism 7, by GraphPad (2 mentions)
Lps ek, by InvivoGen (2 mentions)
Hl 60, by American Type Culture Collection (2 mentions)
Sw480, by American Type Culture Collection (2 mentions)
Smmc 7721, by American Type Culture Collection (2 mentions)
Mcf 7, by American Type Culture Collection (2 mentions)
Dmi3000b, by Leica (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Facscalibur, by BD (2 mentions)
5 bromo 29 deoxyuridine brdu proliferation assay, by Merck Group (2 mentions)
Arginase assay kit, by Merck Group (1 mentions)
Igepal ca 630, by Merck Group (1 mentions)
Goat anti mouse igg hrp, by Santa Cruz Biotechnology (1 mentions)
Anti ar clone 441, by Santa Cruz Biotechnology (1 mentions)
Hrp goat anti rabbit igg, by Vector Laboratories (1 mentions)
91 protocols using multiskan fc plate reader
1
NF-κB Activation Assay for MPs
2
NF-κB Activation Assay for MPs
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RAW-Blue NF-κB cells (Invivogen) were passaged and plated in a 96 well plate at 100k cells/well in 180 μL DMEM containing 10% HIFBS. Cells were incubated at 37°C and 5% CO2 for 24 h. 100 ul of cells were incubated with varying ratios of MPs at 37°C and 5% CO2 for 18 h. After 18 h, 20 μL of the cell supernatant was placed in 180 μL freshly prepared QuantiBlue (Invivogen) solution and incubated at 37°C/5% CO2 for up to 2 h. The plate was analyzed every hour using a Multiskan FC plate reader (Thermo Scientific) and absorbance was measured at 620 nm BCA Assay- This was performed according to manufacturer’s instruction (Thermo Fischer) with some modifications. 100 million beads were incubated with BCA solution and reacted for 30 mins at 60°C then analyzed every hour using a Multiskan FC plate reader (Thermo Scientific) and absorbance was measured at 562 nm and compared to a standard curve of modified MPLA or Pam2 after subtracting a background of maleimide modified MP.
3
Neutrophil Arginase Activity Quantification
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Whole cell lysates of highly purified neutrophils were prepared in 1% Igepal CA-630 (Sigma Aldrich, St Quentin Fallavier, France), 40 mM Tris-HCl, pH 8, 150 mM NaCl, 5 mM EDTA, 5 mM iodoacetamide, 2 mM PMSF, and protease inhibitor mixture (Complete Mini tablet; Roche Applied Scienc, Sigma Aldrich, St Quentin Fallavier, France) at 4 °C for 30 min. The arginase activity level was determined using the Arginase Assay Kit according to the manufacturer’s instructions (Sigma Aldrich, St Quentin Fallavier, France). The absorbance was measured at 450 nm using a Multiskan FC plate reader (ThermoFisher Scientific, Courtaboeuf, France).
4
Western Blot and Cell Viability Assays
5
Quantifying FGF7, BMP6, and IGF-1 Proteins
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To determine FGF7, BMP6, and IGF-1 protein concentrations in culture media, the hFGF7, hBMP6, and hIGF-1 Quantikine ELISA kits (R&D, DKG00, DY507, DG100B, Minneapolis, Minnesota, United States) were used according to the manufacturer's protocol. Plates were measured on a ThermoScientific Multiskan FC plate reader (ThermoScientific Multiskan FC, Waltham, MA, USA).
6
Blue Light Stimulation Assay
7
SARS-CoV-2 RBD Protein Binding Assay
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RBD proteins of SARS-CoV-2, RaTG13 GX-pangolin, GD-pangolin, and SARS-CoV (1 μg/mL) were coated on a MaxiSorp Nunc-immuno 96-well plate (Thermo Scientific, USA) overnight at 4 °C. Wells were blocked with 5% non-fat milk (Biofroxx, Germany) in PBS for 1 h at room temperature, followed by incubation with serially diluted heat-inactivated sera or mAb in disruption buffer (PBS, 5% FBS, 2% bovine serum albumin (BSA), and 1% Tween-20) for 1 h at room temperature. A 1:2500 dilution of horseradish peroxidase-conjugated goat anti-human IgG antibody (Jackson Immuno Research Laboratories, USA) was added for 1 h at room temperature. Wells were washed five times between each step with 0.2% Tween-20 in PBS. Wells were developed using ABST (Thermo Scientific, USA) for 30 min and read at 405 nm on a Multiskan FC plate reader (Thermo Scientific, USA).
8
Cell Proliferation Quantification via CCK-8
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Cell proliferation was evaluated using the Cell Counting Kit-8 (CCK-8; Dojindo, Kumamoto, Japan), following the manufacturer's instructions. After 48 hours of transfection, the MKN-45 or MGC-803 cells were seeded in a 96-well plate. The analysis was performed with 3 replicates for each group. To each well, 10 μL of CCK-8 solution was added and the cells were incubated at 37°C for 1 hour. The absorbance of the mixture was measured at 450 nm using a Multiskan FC plate reader (51119100, Thermo Scientific, Waltham, MA, USA). The proliferation of cells was determined every 24 hours.
9
Cytotoxicity Screening of Cancer Cell Lines
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Five human cancer cell lines, myeloid leukemia HL-60, hepatocellular carcinoma SMMC7721, lung cancer A-549 cells, breast cancer MCF-7, and colon cancer SW480, were used in the cytotoxic assay. All the cells were cultured in RPMI-1640 or DMEM medium (Hyclone, USA), supplemented with 10% fetal bovine serum (Hyclone, USA).The cytotoxicassay was performed according to the MTS method in 96-well microplates. Briefly, adherent cells (100 μL) was seeded into each well of 96-well cell culture plates and allowed to adhere for 24 h before drug addition, while suspended cells were seeded just before drug addition, each tumor cell line was exposed to the test compound dissolved in DMSO in triplicates for 48 h at 37 °C, with DDP and taxol (Sigma, USA) as positive controls. After the incubation, 20 μL MTS and 100 μL medium was added to each well after removal of 100 μL medium, and the incubation continued for 4 h at 37 °C. The optical density was measured at 492 nm using a Multiskan FC plate reader (Thermo Scientific, USA).
10
Tubulin Polymerization Kinetics
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Tubulin polymerization was induced in a solution of 5 mg/mL porcine tubulin (T240, Cytoskeleton, Inc.), 80 mM piperazine-N,N′-bis(2-ethanesulfonic acid) (PIPES), 1 mM ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid, 2 mM MgCl2, 10% glycerol, 1 mM GTP, and 0, 1, 5, or 10 mM sodium pyrophosphate tetrabasic decahydrate (Sigma). The 20 µL reaction mixture was transferred into a 384 microplate (#242757, Thermo Scientific), and the absorption at 350 nm was measured every 30 s for 40 min at 37°C using a Multiskan FC plate reader (Thermo Scientific).
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