Dexamethasone, ascorbic acid, β-sodium glycerophosphate and inhibitor PD98059, MTT (both from Sigma-Aldrich; Merck KGaA), microtiter plate reader (Bio-Tek Instruments, Inc.), ALP activity assay kit (Nanjing Jiancheng Bioengineering Institute), Bio-Rad protein assay reagent (Bio-Rad Laboratories, Inc.), chemiluminescence detection system (Cell Signaling Technology) and image analysis system (NIH Image, version 1.61).
Microtiter plate reader
Manufactured by Agilent Technologies
Sourced in United States, France
The Microtiter plate reader is a laboratory instrument designed to measure the absorbance, fluorescence, or luminescence of samples in a microtiter plate format. It is used to analyze the optical properties of a wide range of samples, including cell-based assays, enzyme-linked immunosorbent assays (ELISAs), and other biochemical and molecular biology applications.
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Lab products found in correlation
Methyl thiazolyl tetrazolium (mtt), by Merck Group (4 mentions)
Gpo trinder triglyceride assay kit, by Applygen (4 mentions)
Bovine serum albumin (bsa), by Merck Group (4 mentions)
Bca protein assay, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Prism 7, by GraphPad (3 mentions)
96 deep well plate, by Corning (3 mentions)
Celltiter 96 aqueous one solution mts reagent, by Promega (2 mentions)
1 step ultra tmb elisa substrate solution, by Thermo Fisher Scientific (2 mentions)
Recombinant sars cov 2 s protein, by Thermo Fisher Scientific (2 mentions)
Hrp conjugated anti hamster igg h l antibody, by Southern Biotech (2 mentions)
Goat anti mouse igg, by Southern Biotech (2 mentions)
Nunc maxisorp, by Thermo Fisher Scientific (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Corning (2 mentions)
96 well plate, by Corning (2 mentions)
Microtiter plate, by Thermo Fisher Scientific (2 mentions)
96 well high binding microplates, by Corning (2 mentions)
Na star influenza neuraminidase inhibitor resistance detection kit, by Thermo Fisher Scientific (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Agilent Technologies (2 mentions)
172 protocols using microtiter plate reader
1
Osteogenic Differentiation Evaluation
2
Sclerostin's Effect on Cell Proliferation
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Cell proliferation was measured using the RealTime GloTM assay (G9711, Promega, Madison, WI, USA). The assay was performed according to the manufacturer’s instructions and the luciferase reaction was measured every 24 h. To analyze the influence of recombinant human (rh)-sclerostin (100-49, Peprotech, Hamburg, Germany) on cell proliferation, 1500 cells were seeded per well of a 96-well plate and four different concentrations of sclerostin (0; 1; 5; 10 ng/mL) were tested. To achieve a critical number of technical replicates in each independent experiment, six wells were seeded per concentration. After initial seeding, proliferation was measured every 24 h. The medium was completely changed before each measurement and new sclerostin was added with each medium change. Reduction in luciferase substrate was measured using a microtiter plate reader (Biotek, Winooski, VT, USA). The proliferation assay was repeated three times and all experiments showed the same trend. However, due to the large variance, statistical analysis over all three experiments was not sufficient. Therefore, a representative data set is shown in the results.
3
Cell Proliferation Assay using MTS
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Cell proliferation (MTS) assay was performed as described elsewhere [25 (link)]. In brief, we seeded all cells at 4000 cells/well (0.1 ml) in 96-well plates. All cells were incubated overnight at 37 °C for 5–6 days. At each indicated time point, cells were incubated with 20 μl of CellTiter 96 AQueous One solution reagent MTS (Promega) in 100 μl of RPMI-1640 for one hour. Cell number was then estimated using a microtiter plate reader (Bio-tek).
4
Biofilm Biomass Quantification on Contact Lenses
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Biofilm biomass was evaluated by the colorimetric microtiter plate assay as described by O’Toole et al (2011).26 (link) Overnight bacterial biofilms were cultured on CLs of different materials, including HEMA, Ocufilcon D, Filcon II 3, Nesofilcon A, and printed HEMA (colored CLs), which were aseptically placed in 24-well plates and incubated at 37°C for 24 h. After incubation, the CLs were transferred to new plates. The CLs were gently washed twice with sterile phosphate buffered saline (pH 7.3) and air-dried. An aliquot of 200 μL of 0.1% crystal violet solution was added to each well to stain the bacterial biofilm for 15 min at room temperature. Any excess stain was removed by rinsing with distilled water. The biofilm biomass on the CLs was determined by decolorization with 200 μL of 33% acetic acid for 15 min and quantified by measuring the OD at 570 nm using a microtiter plate reader (BioTek Instruments, Inc., USA). All experiments were performed in triplicate and repeated three times.
5
Quantification of Hepatic Triglycerides
6
Microtiter Plate Biofilm Quantification
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A 96-well
microtiter plate was prepared according to the method described above.
The plate was incubated at 37 °C for 16 h under static conditions. Trans-Chalcone (20 μg/mL) was used as a positive control.23 (link),24 (link) After incubation, non-adherent cells were removed, and the wells
were washed twice using PBS. The plate was dried for 15 min in laminar
airflow, and the biofilm was stained with filtered 0.1% (w/v) crystal
violet (CV) for 20 min at room temperature. The excess stain was removed
by washing with PBS three times. Then, 95% ethanol was added to the
wells and incubated for 10 min. Absorption of the CV stain was recorded
at 570 nm using a micro-titer plate reader (BioTek, USA). Each sampling
was done in a triplicate manner. Statistical comparisons between groups
were performed by Student’s t-test, where p < 0.05 was considered to be statistically significant.
microtiter plate was prepared according to the method described above.
The plate was incubated at 37 °C for 16 h under static conditions. Trans-Chalcone (20 μg/mL) was used as a positive control.23 (link),24 (link) After incubation, non-adherent cells were removed, and the wells
were washed twice using PBS. The plate was dried for 15 min in laminar
airflow, and the biofilm was stained with filtered 0.1% (w/v) crystal
violet (CV) for 20 min at room temperature. The excess stain was removed
by washing with PBS three times. Then, 95% ethanol was added to the
wells and incubated for 10 min. Absorption of the CV stain was recorded
at 570 nm using a micro-titer plate reader (BioTek, USA). Each sampling
was done in a triplicate manner. Statistical comparisons between groups
were performed by Student’s t-test, where p < 0.05 was considered to be statistically significant.
7
Anti-Biofilm Activity of Pa-MAP 1.9
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Dilutions (1/100) of overnight cultures of E. coli (ATCC O157) and K. pneumoniae (KPC 001825971) grown overnight in LB medium were incubated in basal medium 2 (BM2) [62 mM potassium phosphate buffer (pH 7), 7 mM (NH4)2SO4, 2 mM MgSO4, 10 μM FeSO4, 0.5% glucose], in 96-well cell culture cluster round bottom microplate in the presence of Pa-MAP 1.9 for 24 h at 37 °C. Positive growth control contained only bacteria, and negative control wells contained broth only. Afterwards, planktonic cells were removed and microplate wells were washed twice with deionized water. The attached bacteria remaining were stained with 100 μL of 0.1% (m/v) crystal violet for 20 min. The microplates was rinsed twice with deionized water, air-dried and resolubilized with 110 μL of ethanol 70%. The content of the microplate was transferred to a new microplate, and the MBIC of Pa-MAP 1.9 was accessed at 595 nm in a microtiter plate reader (Bio-Tek Instruments Inc., VT).
8
Planktonic Growth Rates of Tet38-Overexpressing S. aureus
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The planktonic growth rates of 8 tet38-overexpressed S. aureus isolates were determined as described previously (Wijesinghe et al., 2019 (link)), with some modifications. Briefly, 8 tet38-overexpressed isolates and ATCC 29213 standard cell suspensions were prepared by adjusting the turbidity of suspension to 0.5 McFarland standard in sterile saline. Then, 200 μL of each suspension was inoculated in 20 mL sterile LB broth containing fosfomycin in 0, 1/8 MIC, 1/4 MIC, and 1/2 MIC, respectively, for growth at 37°C and 180 rpm for 24 h. The growth rate of the planktonic bacteria was determined by measuring the optical density (OD) of the suspension in each well of the 96-well plate at 600 nm at 2-h intervals for 24 h using a microtiter plate reader (BioTek, United States). The growth curve was generated in triplicate for each experiment. ATCC 29213 served as the control strain.
9
DPPH Radical Scavenging Assay for Selenium Nanoparticles
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The BSeNPs (10, 20, 30, 40, 50, and 60 µg/mL) were tested for their ability to scavenge DPPH (Abd Elkader et al., 2022 (link)). First, 50 µL of BSeNPs was added to 50 µL of DPPH; the mix was loaded in a microtiter plate and left for 30 min; the generating color was read at 517 nm using a microtiter plate reader (BioTek, Vermont), and the obtained values were applied in the equation
10
Dextran Permeability Assay Protocol
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The permeability study was performed for all groups using a 4 kDa fluorescein isothiocyanate (FITC) Dextran marker (Sigma-Aldrich, CAT:46944-100MG-F). Dextran permeability assay was performed for samples at Day 28. Dextran (10 mg ml−1) was applied to the apical part of Transwell inserts for cell-laden and blank samples and then all inserts were incubated in the incubator. Media was collected from the basal compartment every 60 min and loaded into 96-well plates (Greiner Bio) for 3 h. Using a microtiter plate reader (BioTek Instruments Inc.), Dextran content in the base samples were calculated using a 492 nm-excitation wavelength.
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