Isatis root was purchased from Beijing Shengtaier Biological Technology Co., Ltd. (Beijing, China). The preparation of IRPS from Isatis root was performed by the phenol-sulfuric acid method. Briefly, Isatis root crude extracts were immersed in deionization water for dissolution. Subsequently, 1 mL solution was mixed with 1.0 mL of 6% phenol solution. After being mixed well, 5.0 mL of concentrated sulfuric acid was added and mixed immediately. The solvent was heated at 70 °C for 20 min and stored at 16 °C. The absorbance values of OD570 were measured using an automated plate reader (Bio-Rad, Hercules, CA, USA). All samples were prepared in triplicate. The final concentration of Isatis root polysaccharide was 0.447 mg/mL.
Automated plate reader
Manufactured by Bio-Rad
Sourced in United States
The Automated Plate Reader is a versatile laboratory instrument designed to perform rapid and accurate detection and quantification of various analytes in microplate formats. It utilizes advanced optical technologies to measure absorbance, fluorescence, and luminescence signals, enabling a wide range of applications in areas such as biochemistry, cell biology, and drug discovery.
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Lab products found in correlation
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
Cck 8 assay, by Dojindo Laboratories (1 mentions)
Cck 8 reagent, by Roche (1 mentions)
Celltiter 96 aqueous one solution reagent, by Promega (1 mentions)
Hrp conjugated goat anti mouse igg, by Merck Group (1 mentions)
Microtiter plate, by Corning (1 mentions)
Cytotox 96 assay, by Promega (1 mentions)
Cck 8 assay, by Beyotime (1 mentions)
Cck 8 reagent, by Dojindo Laboratories (1 mentions)
96 well cell culture plate, by NEST Biotechnology (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
13 protocols using automated plate reader
1
Isatis Root Polysaccharide Extraction and Quantification
2
MTT Assay for Cell Viability
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In MTT assay, 2000 cells were planted into 96-well plates for 5 wells in each group and cultured for 72 h. After that, cells were incubated in 100 μl MTT reagent (mixture of 10 μl 5 mg/ml MTT and 90 μl cell culture medium) for 2 h. Then, the MTT reagent was discarded and 100 μl DMSO was added into each well in a dark environment. After 10 minutes, the relative optical density (OD) at 570 nm was measured with an automated plate reader (Bio-Rad, USA).
3
Cell Proliferation Quantification by CCK-8 Assay
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BC cell proliferation was quantified by Cell Counting Kit-8 (CCK-8; Dojindo, Tokyo, Japan) assay in this study. Cells were added to 96-well plates at a density of 3x103/well and incubated for 1 to 5 days. Cell viability was measured using CCK-8 according to the manufacturer’s protocol at an optical density of 450 nm using an automated plate reader (Bio-Rad, USA).
4
Cell Proliferation Assay with CCK-8
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GC cell proliferation was measured with a CCK-8 assay (Dojindo, Tokyo, Japan). Briefly, cells in the logarithmic phase were plated onto 96-well plates at a density of 3000 cells per well. A volume of 10 µL of CCK-8 solution was added to each well at the indicated times (24, 48, 72, and 96 h), followed by 1.5 h of incubation. The relative optical density (OD) was measured at 450 nm using an automated plate reader (Bio-Rad, USA).
5
Cell Viability Assay with PAB Treatment
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After human TE-1, ECA-109, KYSE410, and KYSE520 cells were treated with PAB at various concentrations, cells were seeded in 96-well plates at a density of 1x103 cells/mL and incubated at 24, 48, and 72 hours. Subsequently, 10 μL CCK-8 reagent (Roche Diagnostics, Basel, Switzerland) was added into each single well. Following 1 hour of incubation, the absorbance of CCK-8 was measured by an automated plate reader at 490 nm (Bio-Rad, Hercules, CA, USA).
6
Evaluating EV Impact on Hepatoma Cell Proliferation
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To evaluate the impact of EVs on proliferation of hepatoma cells, the viability of Hep3B cells treated with EVs was determined using an MTS assay. A total of 2.5x103 Hep3B cells were seeded in a 96-well plate and treated with either 1x106 or 3x106 EVs daily for 3 days either with or without differentiated THP-1 cells. Subsequently, 20 µl CellTiter96® AQueous One Solution Reagent (Promega Corporation) was added to each well. Following incubation for 2 h at 37˚C, the reaction was measured using an automated plate reader (Bio-Rad Laboratories, Inc.) at 490 nm. To evaluate the concentration of erlotinib that could be used whilst maintaining the viability of hepatoma cells, MTS assays were used. A total of 2.5x103 Hep3B cells were seeded in a 96-well plate and treated with various concentrations of erlotinib for 3 days. MTS assays were performed in at least duplicate.
7
Epitope Mapping of FMDV 3B Peptides
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Six peptides were synthesized by Genscript Inc. (Nanjin, China) based on the complete amino acid sequence of 3B from FMDV O/CHA/99 (Table 1 ). The purity of these peptides was determined by HPLC to be ≥90%. The peptide ELISA was performed according to the method described by Hohlich et al. [3] (link) to analyze the binding epitopes of different McAbs against 3B. Briefly, microtiter plates (Corning, Salt Lake City, USA) were coated with synthetic peptides. After blocking, hybridoma supernatants containing the McAbs were added to each plate. After washing, HRP-conjugated goat anti-mouse IgG (Sigma, USA) was added and TMB (3, 3′, 5, 5′-tetramethyl-benzidine) substrate (SurModics, USA) was used for color development. The optical density (OD) was measured at 450 nm using an automated plate reader (Bio-Rad, USA).
8
Cytotoxicity Assay for Dendritic Cells
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The lactate dehydrogenase (LDH)-based CytoTox 96 Assay (Promega) was used to determine in vitro cytotoxicity according to the manufacturer’s instructions. Briefly, the culture medium was changed to Opti-MEM supplied with 3 % heat-inactivated FCS for experimental cells immediately before the experiment following the extensive washing to avoid contamination with LDH from FCS. 4 × 104 DCs (target cells)/sample, in quadruplicate were co-cultured with 2 × 105 isolated CD56pos cells (min 90 % purity). Alternatively: 1) DCs were preincubated with anti-DC-SIGN mAbs (to 30 μg/ml) for 30 min and cytotoxicity was performed in the medium containing these antibodies; 2) CD56pos cells were preincubated for 4 h with the C3d peptide blocking NCAM homotypic interaction. After 4 h of incubation for each different experimental condition, released LDH into the culture supernatants was measured with a 30-min coupled enzymatic assay, which results in the conversion of a tetrazolium salt into a red formazan product that is read at 490 nm in an automated plate reader (Bio-Rad).
9
Cell Viability Assessment of SM and Fe3O4-SM
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The cells were seeded into 96-well plates at a density of 1x104 cells/well, prior to being treated with SM or Fe3O4-SM for 16 h. The final concentrations of SM were 2.4, 4.8 and 9.6 µM. Cell viability was determined using a CCK-8 assay (Beyotime Institute of Biotechnology, Haimen, China). Absorbance was read on an automated plate reader (Bio-Rad Laboratories, Inc., Hercules, CA, USA) at 450 nm. Cell growth inhibition is presented as the percentage of untreated controls. Growth inhibition was also determined when the cells were treated with SM or Fe3O4-SM (final concentration of SM, 4.8 µM) for 12, 18, 24 and 48 h. All determinations were performed in triplicate.
10
Cell Proliferation Assay of Gastric Cancer Cells
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A CCK-8 assay was used to determine the cell proliferation ability of the MGC803, MGC803-AR, MKN45 and MKN45-AR cells. In brief, the cells were seeded into 96-well cell culture plates (Nest Biotechnology Co., Ltd., Wuxi, China) at a density of 5×103 cells/well in 100 µl RPMI medium containing 10% FBS. Following culturing in RPMI medium for 24 h at 37°C, apatinib and triptolide were administrated for 24 h. Subsequently, 10 µl of CCK-8 reagent (Dojindo, Inc., Tokyo, Japan) was added to each well and the cells were incubated for 2 h at 37°C according to the manufacturer's protocol. Subsequently, the absorbance was read at a wavelength of 450 nm using an automated plate reader (Bio-Rad Laboratories, Inc., Hercules, CA, USA). The experiments were performed in triplicate.
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