Cytotoxic effects of BSA NPs on HL-60 cells were measured by Cell Counting Kit-8 assay. HL-60 cells were plated in 96-well plates (Costar, Corning, NY) at 5000 to 10,000 cells per well. After incubation for 24 hours, the culture medium was removed, and a complete medium with various concentrations of free DOX, BSA, and DOX-conjugated BSA NPs were used to incubate cells for 24 hours, respectively. Ten microliters of the solution cell proliferation reagent (Promega, Madison, WI) per well was added. Then, the cell viability was measured by a Synergy Neo fluorescence plate reader (BioTek, Winooski, VT) at 490 nm.
Synergy neo fluorescence plate reader
Manufactured by Agilent Technologies
The Synergy Neo fluorescence plate reader is a high-performance instrument designed for a variety of fluorescence-based applications. It features a flexible monochromator-based optical system, high-sensitivity detection, and a temperature-controlled incubator. The Synergy Neo provides accurate and reliable data for researchers working in fields such as cell-based assays, biochemical assays, and molecular biology.
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Lab products found in correlation
3 protocols using synergy neo fluorescence plate reader
1
Cytotoxicity of BSA NPs on HL-60 Cells
2
Fluorescent Peptide Binding Assay
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An H3K9ac14ac18ac peptide (amino acids 1–20) functionalized with C-terminal 5-carboxyfluorescin (FAM) was synthesized by the UNC High Throughput Peptide Synthesis Core facility. Three micrograms of each antibody were serially diluted in a black 384 well plate (Corning #3575) and 10 nM FAM peptide in 25 mM HEPES pH 7.5, 100 mM NaCl, 0.05% NP-40 were added. Polarization was measured on a Synergy Neo fluorescence plate reader (Biotek) with a 485±10 nm excitation filter and a 528±10 nm emission filter. Measurements were scaled to the lowest dilution of protein with a requested polarization of 20 milli-polarization units (mP). Anisotropy units (A) were calculated using the equation A = (2P)/(3-P).
3
Fluorescent Peptide Binding Assay
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Peptides functionalized with N-terminal 5-carboxyfluorescin (FAM) were synthesized by Genscript. All 7-mer motifs were synthesized with flanking glycines to mimic the Kme-OPL design. Binding assays were done in black 384 well plates (Corning #3575). Protein was serially diluted with 10 nM FAM peptide in FP assay buffer (25 mM HEPES pH 7.5, 100 mM NaCl, 0.05% NP-40). Polarization was measured on a Synergy Neo fluorescence plate reader (Biotek) with a 485 ± 10 nm excitation filter and a 528 ± 10 nm emission filter. Measurements were scaled to the last dilution of protein with a requested polarization of 20 milli-polarization units (mP). Anisotropy units (A) were calculated using the equation A = (2P)/(3-P). Dissociation constants were determined by non-linear regression analysis of anisotropy curves by specific binding with Hill slope in GraphPad version 8.3.0.
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