In 96-well plates, after 24 h transfection, cells were maintained in 10% FBS-supplemented DMEM for 0, 24, 48, or 72 h. At every time point, 10 μl of the CCK-8 solution (Dojindo Laboratories, Kumamoto, Japan) was added into each well, and the cells were incubated further at 37°C for 2 h. Optical density was measured at 450 nm wavelength on a Sunrise™ microplate reader (Tecan Group, Ltd., Mannedorf, Switzerland).
Sunrise microplate reader
Manufactured by Tecan
Sourced in Switzerland, Austria, Germany, United States, Japan
The Sunrise microplate reader is a versatile and reliable instrument designed for performing various absorbance-based assays in a microplate format. It is capable of measuring absorbance at multiple wavelengths, enabling a wide range of applications in the field of life sciences research and diagnostics.
Automatically generated - may contain errors
Lab products found in correlation
Methyl thiazolyl tetrazolium (mtt), by Merck Group (9 mentions)
Magellan software, by Tecan (8 mentions)
Cck 8 solution, by Dojindo Laboratories (6 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (6 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (5 mentions)
Mtt reagent, by Merck Group (5 mentions)
Cell counting kit, by 7Sea Biotech (4 mentions)
Cck 8, by Dojindo Laboratories (4 mentions)
Bovine serum albumin (bsa), by Merck Group (3 mentions)
Prism 7, by GraphPad (3 mentions)
Mtt solution, by Merck Group (3 mentions)
Agilent 1100, by Agilent Technologies (3 mentions)
Tmb substrate set, by BioLegend (3 mentions)
G1049, by Merck Group (3 mentions)
Ab208348, by Abcam (2 mentions)
Matrigel, by BD (2 mentions)
Thiazolyl blue tetrazolium bromide, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Magellan 7 x 2010, by Tecan (2 mentions)
Sterile 96 well polystyrene plates, by Sarstedt (2 mentions)
355 protocols using sunrise microplate reader
1
Cell Proliferation Assay Using CCK-8
2
Plasma BNP Determination in Newborn Rats
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Blood (~2 ml) was collected from 10newborn rats per group following decapitation, and placed into an EDTA anticoagulation tube. Blood samples were centrifuged at 1,600 × g for 20 min at 4°C, and plasma was removed and stored at −80°C for further study. Plasma BNP concentrations were measured using the Rat BNP 32 ELISA kit (cat. no. ab108815; Abcam), according to the manufacturer's protocol. The plates were read at a wavelength of 450 nm (Tecan Sunrise Microplate reader; Tecan Group Ltd., Männedorf, Switzerland) and the level of BNP was determined for each sample from a standard curve. In the present study, 10 samples from each group were used and experiments on each sample were repeated 3 times.
3
Quantitative Serum IgE and IgG2a Assay
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Serum total IgE and IgG2a levels were measured using OptEIA Mouse kits (BD Pharmingen, San Diego, CA, USA), according to the manufacturer’s instructions. The assays were quantified based on optical density (450 nm), which was read using a calibrated Sunrise microplate reader (Tecan Group, Kawasaki, Japan) and XFluor4 software (Tecan Group).
4
Quantifying sFlt-1 and PLFG in Rat Placenta and Serum
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Rat soluble fms-like tyrosine kinase-1 (sFlt-1, #MBS2602003) and placenta growth factor (PLFG, #MBS026910) were tested in placenta and serum with enzyme-linked immunosorbent assay (ELISA) according to the manufacturer’s instruction (Mybiosource). In brief, 1-mg protein of rat placenta (homogenized in Pierce™ IP lysis buffer containing 1% (v/v) protease inhibitor cocktail) and 100-μl serum were added as samples. Samples and standard were then covered and incubated at room temperature for 90 min with gentle shaking. After washing twice, biotinylated antibodies were added and incubated at room temperature for another 60 min. After washing for three times, HRP-Avidin was added and incubated for 30 min at room temperature with gentle shaking. After washing for five times, color reagent was added in dark with gentle shaking. After 30 min of incubation, stop solution was added and the absorption was read at 450 nm immediately with Sunrise™ microplate reader (Tecan Group) and analyzed by Magellan™ software (Tecan Group).
5
Cell Viability Measurement After Transfection
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Cells were cultured for 6, 12, 24 and 48 h after transfection and cell viability was measured using Cell Counting Kit-8 (CCK-8) according to the manufacturer’s instructions (Solarbio, Beijing, China). The Sunrise microplate reader (Tecan Group, Männedorf, Switzerland) measured the absorbance at 450 nm.
6
Cell Viability Assay with CCK-8
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Cell viability was detected using a CCK-8 kit (cat. no. CK04; Dojindo Molecular Technologies, Inc.) according to the manufacturer's protocol. Transfected SK-Hep1 and SNU-387 cells (1×104 cell/well) were cultured in 96-well plates containing DMEM at 37°C with 5% CO2 for 0, 24 and 48 h. CCK-8 solution (10 ml) was added into the cells and incubated for another 4 h. A micro-plate reader (Sunrise Microplate Reader; Tecan Group, Ltd.) was used to detect the absorbance at a wavelength of 450 nm.
7
Cell Proliferation and Colony Formation Assays
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Transfected cells were harvested at 24 h posttransfection and used for evaluating cell proliferation. For the CCK-8 assay, cells were seeded in 96-well plates at a density of 2 × 103 cells per well. At 0, 24, 48, and 72 h after seeding, the CCK-8 assay was conducted by adding 10 μL of the CCK-8 solution (Dojindo Molecular Technologies, Inc., Kumamoto, Japan) into each well. Following 2 h of incubation at 37°C, the absorbance was measured at 490 nm using a SUNRISE Microplate Reader (Tecan Group, Ltd., Mannedorf, Switzerland).
For the colony formation assay, 1 × 103 transfected cells per well were seeded in 6-well plates. Next, the cells were incubated at 37°C in the humidified chamber comprising 5% CO2 for 2 weeks. On day 15, the cells were fixed in 4% polyformaldehyde (Sigma‐Aldrich, St. Louis, MO) and rinsed twice with phosphate-buffered saline (PBS). The colonies were then counted (≥50 cells) under a light microscope (Olympus Corporation, Tokyo, Japan).
For the colony formation assay, 1 × 103 transfected cells per well were seeded in 6-well plates. Next, the cells were incubated at 37°C in the humidified chamber comprising 5% CO2 for 2 weeks. On day 15, the cells were fixed in 4% polyformaldehyde (Sigma‐Aldrich, St. Louis, MO) and rinsed twice with phosphate-buffered saline (PBS). The colonies were then counted (≥50 cells) under a light microscope (Olympus Corporation, Tokyo, Japan).
8
Measuring Atrial Fibroblast Proliferation
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The proliferation of atrial fibroblasts isolated from Ang‐II‐infused mice was measured with the Cell Counting Kit‐8 (CCK‐8; Dojindo Molecular Technologies, Kumamoto, Japan) according to the manufacturer's instructions.30 Atrial fibroblasts were plated in 96‐well plates, serum‐starved for 24 h and exposed to Ang‐II, DB1976, siRNA‐Smad3 or Ad‐PU.1 according to our assays. Following incubation, CCK‐8 solution was added for 4 h and the absorbance at 450 nm (OD450) was read with a Sunrise microplate reader (Tecan Group, Ltd., Männedorf, Switzerland).
9
Evaluating hLEC Viability with KAL
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hLECs were seeded in 96-well plates at a cell density of 5,000 cells per well and were allowed to attach overnight. The cells were treated with Ad-KAL/Ad-GFP or rKAL/PBS for 12–48 h. Cell viability was measured using CCK8 assay (Dojindo Molecular Technologies, Inc., Kumamoto, Japan). The absorbance value at 450 nm was read using a Sunrise Microplate Reader (Tecan Group Ltd., Männedorf, Switzerland).
10
Rat LEC Viability Assay
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Rat LECs were seeded in 96-well plates (Corning) at a cell density of 1 × 104 cells per well, and were allowed to attach overnight. The cells were treated with DMEM (control group) or M2b macrophage supernatant (M2b group) for 24 h. Cell viability was then measured using the CCK-8 assay (Beyotime Biotechnology, Shanghai, China). The absorbance value at 450 nm was read using a Sunrise Microplate Reader (Tecan Group Ltd., Männedorf, Switzerland).
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