HCT116 cells were seeded on cell‐culture dishes coated with polyethyleneimine and transfected with siRNAs. At 24 h after transfection, cells were transferred to ultra‐low cluster plates (Corning, NY, USA). After a further 24 h, cells were treated with 1 μg /mL ADR for 48 h and subjected to ATP measurement assay using Cell Titer‐Glo Luminescent Viability Assay (Promega) according to the manufacturer's protocol. The fluorescence of the solution was measured by an ARVO X3 plate reader (Perkin Elmer, Waltham, MA, USA) according to the manufacturer's protocol.
Arvo x3 plate reader
Manufactured by PerkinElmer
Sourced in United States
The ARVO X3 plate reader is a high-performance microplate reader designed for efficient and accurate multi-mode detection. It offers a range of detection modes, including absorbance, fluorescence, and luminescence, enabling comprehensive analysis of various samples and assays.
Automatically generated - may contain errors
Lab products found in correlation
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Celltiter glo luminescent viability assay, by Promega (1 mentions)
Ultra low cluster plate, by Corning (1 mentions)
Prestoblue cell viability reagent, by Thermo Fisher Scientific (1 mentions)
Phosphorus standard solution, by Fujifilm (1 mentions)
Biotek synergy h1, by Agilent Technologies (1 mentions)
Celltiter glo luminescent cell viability assay, by Promega (1 mentions)
5 protocols using arvo x3 plate reader
1
Evaluating Doxorubicin Cytotoxicity in HCT116 Cells
2
Cathepsin L Activity Assay for Cell Lines
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Cathepsin L activity assay was carried out using an Cathepsin L Activity Assay Kit (Promokine, Heidelberg, Germany) following the manufacturer's protocol. Briefly, HEK293T cells transfected with cystatin C expression plasmid or mock plasmid were collected at 36 h after transfection. HCT116 p53+/+ or p53−/− cells treated with 2 μg/mL ADR for 2 h were collected 48 h after treatment. Each siRNA was transfected 24 h before ADR treatment. The cells were lysed with lysis buffer provided in the kit. Protein concentration of each lysate was measured by Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA) and diluted to 1.20 μg/μL. After adding reaction buffer and fluorescent substrate to the lysates, samples were incubated for 1 h at 37°C, and fluorescence was measured at ex 405nm/em 500nm in the ARVO X3 plate reader (Perkin Elmer).
3
Monitoring Cell Growth and Viability
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To monitor cell growth, the Presto-Blue Cell Viability Reagent (Thermo Fisher) was used according to the manufacturer’s protocol. Briefly, 1 × 104 cells / 150 µl / well of U2OS cells that stably express the RAD9 gene were cultured in 96-well dishes with hydroxyurea. Every 24 hr, the 96 well dish was monitored for cell growth. Presto-Blue (17 µl) was added to each well, followed by an incubation in a CO2 incubator at 37 ˚C for 10 min. Subsequently, 75 µl of 10% SDS was added to stop the reaction, and the plate was placed in an ARVO-X3 plate reader (Perkin Elmer). A 531 nm excitation filter and a 616 nm emission filter were used, and for each cell line and hydroxyurea condition, triplicate samples were measured and the average values were plotted in the graph. To monitor the cell viability upon hydroxyurea exposure, a colony formation assay was performed. First, U2OS cells that express the RAD9 protein were treated with 0.2 mM hydroxyurea for 36 hr. The cells were collected by trypsinization, and 500 cells / 10 ml were plated onto 10 cm dishes, followed by an incubation at 37 ˚C in a CO2 incubator for 11 days. The plates were stained with crystal violet/methanol.
4
Quantifying Inorganic Phosphate in Plants
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Inorganic phosphate (Pi) contents in soybean leaves and Arabidopsis seedlings were examined using a molybdate colorimetric assay73 (link),74 (link). Frozen powdered soybean leaf samples used for gene expression analysis were also used to analyze Pi content. For Arabidopsis, three seedlings from each pot were collected in 2-ml tubes, immediately weighed to determine fresh weight, and frozen in liquid nitrogen. A total of 20–45 mg of each sample was mixed with 1%, v/v, acetic acid (20 µl/mg sample weight), vortexed for 15 min, and incubated at 42 °C for 1 h in a heat block. Following centrifugation at 18,300 × g for 5 min at room temperature, the supernatant was used for Pi assay. Reaction solutions containing 140 µl molybdate solution (master mix of 6:1 0.42%, w/v, ammonium molybdate in 1 N H2SO4 and 10%, w/v, ascorbic acid in water) and 60 µl supernatant were incubated in a 96-well plate at 42 °C for 20 min using a thermal cycler. The absorbance of 100 µl of the reaction solutions at 820 nm was measured using an ARVOX3 plate reader (PerkinElmer, Waltham, MA, USA) or a BioTek Synergy H1 (BioTek Instruments Inc., Winooski, VT, USA). The amount of Pi in the solution was calculated using a calibration curve based on a diluted phosphorus standard solution (FUJIFILM Wako, Osaka, Japan) or KH2PO4.
5
Cell Proliferation Assay with Akt Inhibitor
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For the proliferation assay and Akt inhibitor experiments, the cell numbers were evaluated by the CellTiter-Glo Luminescent Cell Viability Assay (Promega) at the indicated times. The luminescence of cell lysates was measured by an ARVO X3 plate reader (PerkinElmer) according to the manufacturer’s protocol.
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