Serum was isolated from mice by centrifugation after allowing whole blood to clot at room temperature. ELISA was performed on diluted samples using the mouse CCL2 (MCP-1) Ready-SET-Go! ELISA reagent set (eBioscience) according to manufacturer's protocol. Absorbance values were measured by the Dynex MRX-TC Revelation microplate reader/colorimeter (Dynex Technologies, Chantilly, VA) at 450 nM wavelength. Protein concentrations were determined with reference to a serial dilution of recombinant mouse MCP-1 protein (eBioscience). Linear equation for the standard curve and sample analyses was generated by exporting results to Microsoft Excel (version 2010).
Mrx tc revelation
Manufactured by Dynex
Sourced in United States
The Dynex MRX-TC Revelation is a laboratory instrument designed for performing microplate-based assays. It features temperature control capabilities to maintain samples at desired temperatures during testing.
Automatically generated - may contain errors
Lab products found in correlation
Aqueous one solution cell proliferation assay, by Promega (1 mentions)
Tmb solution, by Merck Group (1 mentions)
Mouse iga elisa kit, by Fortis Life Sciences (1 mentions)
Dulbecco s modified eagle medium, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
9 protocols using mrx tc revelation
1
Measuring Serum MCP-1 Levels in Mice
2
Curcumin Modulates Cell Viability
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Cell viability was measured using the AQueous One Solution cell proliferation assay according to the manufacturer's protocol (Promega Corporation, Madison, WI). Cells were treated with the indicated concentrations of curcumin and compared to DMSO vehicle controls. Absorbance values at 490 nM wavelength were recorded with the Dynex MRX-TC Revelation microplate reader/colorimeter (Dynex Technologies). The absorbance value without curcumin treatment was designated +1 in RAW 264.7 cells and the relative fold change in viability was determined for various treatments after normalizing to background absorbance levels. Absorbance values at baseline were designated +1 for THP-1 cells and primary human macrophages.
3
Microdilution Susceptibility Testing of FAR
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Susceptibility testing was carried out using the microdilution method according to the EUCAST protocol (version 12.0, 2022) [37 ]. Overnight cultures of S. aureus were washed twice in PBS and adjusted to 5 × 105 bacteria/mL in MHB. The stock solution of FAR (75 mM, Sigma-Aldrich, Steinheim, Germany), prepared in 96% ethanol (Centralchem, Banská Bystrica, Slovakia), was diluted in MHB medium to obtain final concentrations in the wells, namely 1000, 500, 250, 125 and 62.5 µM (corresponding to 222, 111, 55.5, 27.75, 13.88 mg/mL). Next, 100 μL of the bacterial cell suspension and 100 μL of FAR at the appropriate concentration were added to the wells. Bacteria without treatment served as a positive control. From each sample, three parallel wells were prepared. The plates were incubated statically for 24 h at 37 °C. The effectiveness of FAR was determined in terms of MIC50. The intensity of bacterial growth was measured spectrophotometrically at OD570 (Dynex MRX-TC Revelation, Dynex Technologies, Chantilly, VA, USA) against the control with MHB.
4
Serum Urea Nitrogen Quantification
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Serum was collected biweekly using the MaxDiscovery Blood Urea Nitrogen Enzymatic Assay Kit (Bioo Scientific Corporation, Austin, TX) according to manufacturer's protocol. Blood was obtained via submandibular bleeds and serum was isolated by centrifugation after blood clotting. Absorbance values were determined using the Dynex MRX-TC Revelation microplate reader/colorimeter (Dynex Technologies, Chantilly, VA). Results were exported to Microsoft excel (v2013) for analysis.
5
Quantifying Inflammatory Cytokines in Joints
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Serum was isolated and synovial aspirates were collected by rinsing joint spaces with PBS. ELISA was performed on diluted samples using the mouse IL-1β and CXCL1kits (eBioscience) according to manufacturer's protocol. Absorbance values were measured by the Dynex MRX-TC Revelation microplate reader/colorimeter (Dynex Technologies).
6
Antigen-Specific Antibody Titer Assay
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To test the titer of the generated fbAB, the antigen was coated onto high protein binding, polystyrene, 8-well microplate strips (Greiner) at a concentration of 200 ng/well with 0.1 M NaCO3. The strips were incubated for 2 h at 37 °C for coating and then blocked with 2% BSA (in PBS) for 1 h at 37 °C. After washing the strips for 3 times with PBS containing 0.1% Tween 20 (PBS-T), fbAB was added into the wells at the indicated dilutions (prepared with PBS-T) and incubated for 1 h at RT. After washing for 4 times, anti-rabbit IgG-HRP (EpiGentek) (50 µl, 1:2000 dilution in PBS-T) was added and incubated for 30 min at RT. After washing for 4 times, 100 µl of TMB solution (EMD Millipore Corp.) were added per well and blue color development was monitored for 2–10 min. The reaction was stopped with an equal volume of 1 M HCl and the optical density was measured with a microplate reader (MRX-TC Revelation, Dynex Technologies) at a wavelength of 450 nm.
7
Adipogenic Differentiation of eMSCs
8
Quantification of Fecal IgA Levels
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Fecal samples were resuspended in PBS at a concentration of 50 mg/mL by extensive vortexing, allowed to stand for 20 min, and centrifuged at 16,000 g for 10 min to collect supernatant as described (Haneberg et al., 1994 (link)). Each supernatant was assessed for IgA using the mouse IgA ELISA kit with suitable dilutions, according to the manufacturer’s instructions (Bethyl, Montgomery TX), and the absorbance was measured at a wavelength of 450 nm using the Dynex MRX TC Revelation microplate reader (Dynex Technologies, Chantilly VA) (Ruiz et al., 2017 (link)).
9
Evaluating Glass-Ionomer Cytotoxicity on C2C12 Cells
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In order to analyze the cytotoxicity of our experimental glass-ionomer, mouse myoblast cell line C2C12 (American Type Culture Collection) and methyltetrazolium (MTT) assay were utilized. C2C12 cells were cultured in Dulbecco's modified Eagle medium (Sigma-Aldrich) that was supplemented with 100 U/mL penicillin, 100 mg/mL streptomycin (Sigma-Aldrich), and 15% fetal bovine serum (Biocell Laboratories) at 37°C in a humidified atmosphere supplied with 5% CO 2. The prepared glass-ionomer specimens were rinsed with phosphate-buffered saline and minimal essential culture medium (3×) and placed at the bottom of a 6-plate culture dish and 5×10 4 C2C12 cells were cultured in each well (n=6) containing regular culture media for 72 h at 37°C in 5% CO 2. The metabolic activity of cultured cells was assessed using MTT assay, after 72 h of culturing, according to a previously published method 13) (link) . Briefly, 200 µL of MTT was added to each well, and the wells were incubated for 4 h followed by the addition of 200 µL of DMSO. Optical density automated plate reader MRXTC revelation (Dynex Technologies) at 570 nm was utilized to evaluate the percentage of viable cell viability.
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