PARP1 (ThermoFisher), olapBFL and nTLZ or talazoparib (control) were added to wells in a 96 well plate in complete DMEM cell culture media in a total volume of 200 μl. PARP1 and olapBFL were brought to 100 nM. nTLZ was added at a total talazoparib concentration of 100 nM. Control experiments lacked nTLZ or had 100 nM free talazoparib. NTLZ was added immediately before measurements and the solution was mixed briefly prior to measuring. Anisotropy and fluorescence intensity of olapBFL was measured in a plate fluorimeter (Tecan) with excitation at 488 nm and emission collected between 505–515 nm. Measurements were made every 3 minutes for up to 60 minutes at room temperature.
Plate fluorimeter
Manufactured by Tecan
Sourced in Switzerland
The Plate Fluorimeter is a laboratory instrument designed to measure the fluorescence intensity of samples in a multi-well plate format. It provides accurate and sensitive detection of fluorescent signals, enabling researchers to quantify a wide range of biological and chemical analytes in a high-throughput manner.
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Lab products found in correlation
3 protocols using plate fluorimeter
1
PARP1 Inhibitor Binding Kinetics
2
Determining Critical Micelle Concentration
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A duplex serial dilution was prepared in a 96-well plate from 20 µM to 10 nM in 50 μl of Ca2+/Mg2+ free DPBS with Nile Red (0.25 µg), and the plate was agitated at 37 °C in the dark for 2 h. Fluorescence was measured on a plate fluorimeter from Tecan (Mannedorf, Switzerland) at excitation 535 ± 10 nm and emission 612 ± 10 nm. The CMC was defined, as previously described80 (link), as the intersection point on the plot of the two linear regions of the Nile Red fluorescence versus duplex concentration.
3
Nile Red-Based CMC Determination
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A duplex serial dilution was prepared in a 96-well plate from 20 μM to 10 nM in 50 uL of Ca2+/Mg2+ free DPBS with Nile Red (0.25 μg), and the plate was rotated at 37°C in the dark for 2 h. Fluorescence was measured on a plate fluorimeter (Tecan) at excitation 535 ± 10 nm and emission 612 ± 10 nm. The CMC was defined, as previously described 25 , as the intersection point on the plot of the two linear regions of the Nile Red fluorescence versus duplex concentration.
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