Normal PmECs and breast cancer cells were seeded onto 96-well plates at a density of 1 × 105 cells/mL, and left overnight to attach. Culture medium was removed and the cells were treated with serial dilutions of lutein (0.5–2.0 μM) in fresh culture medium with 10% FBS, or cultured in medium with DMSO (in the same concentration as carotenoid samples) as a control. Cell viability was determined after 48 h by MTT assay using CellTiter96® Non-Radioactive Cell Proliferation kits (Promega, Madison, WI, USA), in which the yellow tetrazolium salt [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] is reduced by mitochondrial dehydrogenase in viable cells to purple, insoluble crystals of formazan. Cells were incubated for 4 h with MTT solution at 37 °C and formazan crystals were solubilized in a lysing buffer overnight at room temperature (RT). The product was quantified by measurement of absorbance at a 570 nm wavelength with the use of a BioTek Synergy™ H4 Hybrid Multi-Mode Microplate Reader (BioTek Instruments, Winooski, VT, USA). All experiments were performed in triplicate and yielded similar results. Results are representative of an average of three independent experiments. Data are presented as proportional viability (%) by comparing the treated group with the untreated cells, the viability of which was assumed as 100%.
Biotek synergy h4 hybrid multi mode microplate reader
Manufactured by Agilent Technologies
Sourced in United States
The BioTek Synergy H4 Hybrid Multi-Mode Microplate Reader is a versatile laboratory instrument that can perform various detection modes, including absorbance, fluorescence, and luminescence, within a single platform. The device is designed to accommodate multiple microplate formats and can be used for a wide range of applications in life science research and drug discovery.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using biotek synergy h4 hybrid multi mode microplate reader
1
Lutein Inhibits Breast Cancer Cell Viability
2
Carotenoid Effects on ARPE-19 Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ARPE-19 cells were seeded in 96-well plates at a density of 2 × 103 cell/well and cultured in 100 μL medium overnight for attachment. Then, the culture medium was removed and the cells were treated with serial dilutions of carotenoids (0.5–2.0 μM) in fresh culture medium with 10% FBS or cultured in a medium with DMSO as a control. Cell viability was determined after 48 h by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay using the CellTiter96® Non-Radioactive Cell Proliferation Kits (Promega, Madison, WI, USA), in which the yellow tetrazolium salt is reduced by mitochondrial dehydrogenase of viable cells to purple, insoluble crystals of formazan. Cells were incubated for 4 h with MTT solution at 37 °C. Then, formazan crystals were solubilized in lysing buffer overnight at room temperature (RT). The reaction product was quantified by measurement of absorbance at a 570 nm wavelength using a BioTek Synergy™ H4 Hybrid Multi-Mode Microplate Reader (BioTek Instruments Inc., Winooski, VT, USA). All experiments were performed in triplicate. Results are representative of an average of 3 independent experiments. Data are presented as proportional viability (%), by comparing the treated group with the untreated cells, the viability of which is measured to be 100%.
3
GFPssrA Denaturation Monitoring
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Reaction mixtures were prepared with hydrogenated proteins (70 μM GFPssrA, 20 μM PAN, and 20 μM 20S) in an H2O buffer containing 20 mM Tris-HCl (pH 7.5), 100 mM NaCl, and 10 mM MgCl2. The following conditions were measured: isolated GFP, GFP, and PAN or GFP, PAN, and 20S with 0 mM ATP and 10 mM MgCl2, 10 mM ATP and 20 mM MgCl2, 50 mM ATP and 100 mM MgCl2, and 100 mM ATP and 200 mM MgCl2. The denaturation of GFPssrA was monitored (λex = 400 nm and λem = 509 nm) by using a plate reader spectrophotometer (BioTek Synergy H4 Hybrid Multi-Mode Microplate Reader; BioTek Instruments, Winooski, VT). Fluorescence was measured over 1 h at 55°C with a time resolution of 40 s. Fluorescence curves as a function of time were generated by normalization to the first measurement point, which was defined as 100%.
4
ATP Hydrolysis Kinetics of PAN Proteasome
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The rate of ATP hydrolysis by PAN was measured offline in the same conditions as the SANS experiments. For this purpose, the amount of inorganic phosphate was recorded as a function of time. Reaction mixtures were prepared with hydrogenated proteins (70 μM GFPssrA, 20 μM PAN, and 20 μM 20S) in an H2O buffer containing 20 mM Tris-HCl (pH 7.5), 100 mM NaCl, 100 mM ATP, and 200 mM MgCl2. The following conditions were measured in triplicate: isolated PAN, PAN and GFP, PAN, 20S and GFP, isolated GFP, isolated 20S, and buffer. The samples were incubated at 55°C and monitored over 1 h with a 20 μL reaction mixture measured at each time point. The aliquots were incubated on ice to stop the reaction and diluted 150 times in the buffer. 4 μL were then supplemented with 36 μL of assay buffer and 200 μL of malachite green reagent (MAK113; Sigma-Aldrich). After 30 min of incubation at room temperature, absorbance at 630 nm was analyzed by using a plate reader spectrophotometer (BioTek Synergy H4 Hybrid Multi-Mode Microplate Reader; BioTek Instruments). A standard phosphate curve was used to estimate the amount of phosphate released during the hydrolysis of ATP. Phosphate concentrations were fitted with a biexponential process (Eq. 1 ) in which intensities were replaced by concentrations.
5
Thermal Denaturation of GFPssrA Proteasome
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Reaction mixtures were prepared with hydrogenated proteins (70 μM GFPssrA, 20 μM PAN, and 20 μM 20S) in an H2O working buffer containing 20 mM Tris-HCl (pH 7.5), 100 mM NaCl, 100 mM ATP, and 200 mM MgCl2. The following conditions were measured: isolated GFP, GFP, and PAN and GFP, PAN, and 20S at different temperatures (30, 40, 45, 50, 55, 60, and 65°C). The denaturation of the GFPssrA was monitored (λex = 400 nm and λem = 509 nm) using a plate reader spectrophotometer (BioTek Synergy H4 Hybrid Multi-Mode Microplate Reader; BioTek Instruments). Fluorescence was measured over 1 h at each temperature with a time resolution of 40 s. Fluorescence curves as a function of time were generated by normalization to the first measurement point, which was defined as 100%.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!