The BMSCs transfected with the miR-214 or anti-miR-214 plasmids, or treated with JNK inhibitor or p38 inhibitor were seeded at a density of 2×105 cells/well in 24-well plates and subjected to osteogenic differentiation. The BMSCs were lysed with RIPA lysis buffer (Thermo Fisher Scientific, Wilmington, DE, USA) supplemented with protease inhibitor. The protein concentration was determined by Bradford assay (Bio-Rad, Hercules, CA, USA) on a microplate spectrophotometer (Tecan, Port Melbourne, Australia). ALP activity was determined using an ALP activity kit (Nanjing Jiancheng Biotech, China) on a microplate spectrophotometer (Tecan).
Microplate spectrophotometer
Manufactured by Tecan
Sourced in Switzerland, Austria, Germany, United States, United Kingdom, Australia
The Microplate spectrophotometer is a laboratory instrument designed to measure the absorbance of samples in microplates. It is capable of quantifying various analytes, such as proteins, nucleic acids, and small molecules, by detecting the light absorption of the samples across a specified wavelength range.
Automatically generated - may contain errors
Lab products found in correlation
Bradford assay, by Bio-Rad (3 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (3 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions)
Alp activity kit, by Nanjing Jiancheng (2 mentions)
Cck 8, by Tecan (2 mentions)
Mtt dye, by Merck Group (2 mentions)
Ifn γ, by R&D Systems (2 mentions)
Mtt solution, by Merck Group (2 mentions)
Mtt assay, by Merck Group (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Mts reagent, by Promega (1 mentions)
Tacs annexin 5 fitc apoptosis detection kit, by R&D Systems (1 mentions)
Prism software 8, by GraphPad (1 mentions)
Cck 8, by Beyotime (1 mentions)
Lps from e coli 0111 b4, by Merck Group (1 mentions)
Vemurafenib plx 4032, by Selleck Chemicals (1 mentions)
Cell proliferation reagent wst 1, by Roche (1 mentions)
Dnmt activity assay kit, by Abcam (1 mentions)
75 protocols using microplate spectrophotometer
1
Osteogenic Differentiation of BMSCs
2
Antimicrobial Activity Evaluation of Cath-MH
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All ATCC microorganisms were bought from the Guangdong Institute of Microbiology. The antimicrobial capability of cath-MH was evaluated through minimal inhibitory concentrations (MICs) by the twofold microdilution method as reported previously (Zeng et al., 2018 (link)). In brief, after growing in Muller-Hinton (MH) broth at 37°C to exponential phase, microbes were then diluted with fresh MH broth to reach 106 CFU/ml density. An equal volume of microbial inocula was added into 96-well plates with different concentrations of cath-MH diluted in MH broth. Plates were incubated at 37°C for 14 hr, and the MIC values at which there was no microbial growth were measured at 600 nm with a microplate spectrophotometer (Tecan Trading AG, Männedorf, Switzerland). All experiments were carried out in triplicates with ampicillin and meropenem as positive control.
3
Probe Quenching Efficiency Evaluation
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The probe to be tested and the corresponding probe labeled only with 5'-fluorophore were synthesized by Sangon Biotech Co., Ltd. and diluted to 10 μM. The fluorescence of the probe at 250 nM (the common concentration of probes in RT-qPCR reaction) was measured on a microplate spectrophotometer (Tecan Trading AG, Mannedorf, Switzerland), and the quenching efficiency of the probe was evaluated by comparing the fluorescence intensity with that of the corresponding probe labeled only with 5'-fluorophore.
4
Synergistic Anti-Proliferative Effects of α-Mangostin
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Cell proliferation was measured by the colorimetric 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay (MTT assay; Sigma-Aldrich). We incubated 5×10³ cells/well in 96-well plates and different drug concentrations for 24 h, 48 h and 72 h. To evaluate the synergistic inhibitory effect of α-Mangostin combined with chemotherapeutic drugs on cell proliferation, we used 10 nmol/L α-Mangostin plus different concentrations of chemotherapeutic drugs against melanoma cells. After treatments, MTT 4 h at 37°C, after which supernatant fluids were discarded, and formazan crystal were dissolved in DMSO (150 μL / well). Optical density (OD) values were then read in a microplate spectrophotometer (Tecan Group Ltd., Mannedorf, Switzerland) at 540 nm. The cell proliferation index was calculated according to the following formula: experimental OD value/ control OD value.
5
Quantifying Inflammatory Cytokines Levels
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IL-1β levels were quantified using the Cymax™ Human ultrasensitive IL-1β ELISA assay (L.O.D. 1.90 pg/ml; Ab Frontier, Korea).
IL-6 and TNF-α serum levels were quantified using the Human IL-6 PicoKine ELISA kit (L.O.D.<0.3 pg/ml) and Human TNF-α PicoKine ELISA kit (L.O.D.<1 pg/ml), respectively (Vinci-Biochem srl., Italy).
IL-17 serum levels were assessed by Human IL-17A Platinum Kit ELISA (L.O.D. 1.6 pg/ml; Invitrogen, Thermo Fisher Scientific, Italy).
Reagents and assay procedures were prepared according to the manufacturer's guidelines. The products of the enzymatic reactions were immediately read by a Microplate Spectrophotometer (Tecan Group Ltd., Switzerland) and by a Microplate Reader (BioTek Instruments Inc., USA). A standard curve was created from plate data, and the unknown values were extrapolated in Microsoft Excel.
IL-6 and TNF-α serum levels were quantified using the Human IL-6 PicoKine ELISA kit (L.O.D.<0.3 pg/ml) and Human TNF-α PicoKine ELISA kit (L.O.D.<1 pg/ml), respectively (Vinci-Biochem srl., Italy).
IL-17 serum levels were assessed by Human IL-17A Platinum Kit ELISA (L.O.D. 1.6 pg/ml; Invitrogen, Thermo Fisher Scientific, Italy).
Reagents and assay procedures were prepared according to the manufacturer's guidelines. The products of the enzymatic reactions were immediately read by a Microplate Spectrophotometer (Tecan Group Ltd., Switzerland) and by a Microplate Reader (BioTek Instruments Inc., USA). A standard curve was created from plate data, and the unknown values were extrapolated in Microsoft Excel.
6
Evaluating miRNA-223-3p Effects on LSCC Cell Proliferation
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The effects of miRNA-223-3p expression on the proliferation of LSCC cells were assessed using the Cell Counting Kit-8 (CCK-8; Kumamoto, Japan). Briefly, the cells were transfected for 24 h and plated on 96-well plates. Subsequently, CCK-8 was added to each well at various times and incubated at 37 °C for 1.5 h. The absorbance at 450/630 nM was measured using a microplate spectrophotometer (Tecan Group Ltd., Männedorf, Switzerland). A minimum of five wells were assessed for each group.
7
Cell Viability Assay Using CCK-8
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Macrophages were cultured in 96-well plates with 10,000 cells per well. The CCK-8 assays were performed according to the manufacturer's instructions. After preparing the experimental groups and performing the cell treatments, a 1:10 dilution of the CCK-8 working solution was prepared. Then, 100 µl working solution was removed and added to 96-well plates. The plates were placed in an incubator at 37˚C for 60 min to allow the reaction to occur. After the incubation, absorbance was measured at 450 nm using a microplate spectrophotometer (Tecan Group, Ltd.).
8
FAK siRNA Effects on OS Cell Proliferation
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The effect of FAK siRNA on the proliferation of OS cells was measured using CCK-8 (cat. no. 1166; R&D Systems, Inc., Minneapolis, MN, USA). Cells were seeded into the 96-well plates (10,000 cells per well). CCK8 was used according to the manufacturer's protocol, and 24 h after the transfection of FAK siRNA into the MG-63 cell line, CCK-8 was added into each well and then was incubated at 37°C for 1.5 h. Optical density was measured at 450 nm using a microplate spectrophotometer (Tecan Group Ltd., Zurich, Switzerland).
9
Niloticin cytotoxicity evaluated by MTT assay
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The MTT assay was used to evaluate cell viability.24 (link) The experimental samples were divided into an untreated control group, a DMSO group (treated with 0.5% DMSO), and a niloticin-treated group. Cells were treated with 8, 40, or 100 μg/mL niloticin (six replicates per concentration) for 24 h, and then MTT was added in the medium at 1:10 (v/v; Sigma, St Louis, MO, USA), and the cells were cultured for 4 h. The optical density (OD) was measured using a microplate spectrophotometer at 570 nm (Tecan Group Ltd., Mannedorf, Switzerland).
10
Cell Viability Assay using CCK-8
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Macrophages were cultured in 96 well plates with 10,000 cells per well. The Cell Counting Kit-8 (CCK-8) (Beyotime Institute of Biotechnology) assays were performed according to the manufacturer's instructions. Briefly, after preparing the experimental groups and performing the cell treatments, the CCK-8 working solution was diluted 10-fold and 100 µl was removed and added to 96-well plates. The plates were placed in an incubator (37˚C, 60 min) to allow the reaction to occur. After the incubation, absorbance was measured at 450 nm using a microplate spectrophotometer (Tecan Group, Ltd.).
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