Utilizing F12511’s inherent fluorescence (Mesplet et al., 2005 (link)), F12511 concentration was measured using BioTek Synergy Neo2 Hybrid Multi-Mode Plate Reader (excitation λ: 300nm, emission λ: 380nm). A concentration gradient of F12511 (dissolved in EtOH) ranging from 0–13mM was used to create a standard curve. Nanoparticles were diluted in 1x PBS with volume concentrations ranging from 0–90% (i.e., for 90%, 90μL of nanoparticles and 10μL of 1x PBS are mixed). A total of 100μL was loaded per well and samples were measured in triplicate. The amount of F12511 in Nanoparticle F was determined based on the F12511 concentration gradient trendline equation. To address any possible added fluorescence from the nanoparticles themselves, the control nanoparticles of DSPE-PEG2000/PC were also measured and subtracted from corresponding dilution of Nanoparticle F. This method was used to confirm the amount of F12511 encapsulated.
Synergy neo2 hybrid multi mode plate reader
Manufactured by Agilent Technologies
The Synergy Neo2 Hybrid Multi-Mode Plate Reader is a high-performance microplate reader designed for a wide range of detection modes, including absorbance, fluorescence, luminescence, and time-resolved fluorescence. It offers advanced features and capabilities to support various applications in life science research and drug discovery.
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2 protocols using synergy neo2 hybrid multi mode plate reader
1
Quantifying F12511 Encapsulation in Nanoparticles
2
Senescence Analysis of B-ALL Cells
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Human B-ALL cell lines (106 cells/well in a 12-well plate) were cultured in unconditioned, bone marrow stromal cell-, or adipocyte-conditioned media (depending on the experiment) for 24 h. After the incubation period, cells were harvested and lysates prepared. β-Galactosidase activity was determined using the Senescence β-Galactosidase Activity Assay Kit from Cell Signaling Technology (cat# 23833 S) per the manufacturer’s protocol. Relative Fluorescence Units (RFU) of 4-MU were measured at an excitation wavelength of 360 nm and an emission wavelength of 465 nm using the Synergy Neo2 Hybrid Multi-Mode Plate Reader (BioTek, Winooski, VT).
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