Data analysis software

Manufactured by Horiba

The Data Analysis Software from Horiba is a comprehensive tool designed for the analysis and processing of data generated by various laboratory instruments. It provides a suite of analytical functionalities to aid researchers and scientists in effectively interpreting their experimental results.

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Lab products found in correlation

Deltaflex tcspc system, by Horiba (2 mentions) Fluotime 200, by PicoQuant (1 mentions) Fluorolog fluorimeter, by Horiba (1 mentions) Fluofit software, by PicoQuant (1 mentions) Tcspc, by Horiba (1 mentions)

4 protocols using data analysis software

Fluorescence Lifetime Measurement Protocol

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Fluorescence intensity decays were measured using time-correlated single-photon-counting with a FluoTime200 fluorimeter (PicoQuant, Inc., Berlin, Germany). Excitation was at 295 nm. Emitted light at 340 nm was selected using a monochromator. To avoid effects of Brownian rotation a polarizer oriented at the magical angle was placed on the emission side. A micro channel plate was used to detect the emitted light. The intensity decay data were fitted with FluoFit software (PicoQuant version 4).
Some fluorescence lifetimes were measured using Fluorolog fluorimeter equipped with Nano-LED laser (Horiba Scientific Inc., Edison, NJ). Excitation was at 278 nm. The emitted light at 340 nm was selected using a monochromator. A photomultiplier tube (Hamamatsu 928, Japan) was used to detect emitted light. The intensity decay data were fitted with Data Analysis Software (Horiba Scientific Data Station 2.7).
The data were analyzed in terms of a sum of exponentials: I(t) = Σα_i exp (−t/⊤_i) where α_i are the amplitudes of the intensity decay times ⊤_i with Σα_i = 1.0. The intensity mean lifetime is defined as ⊤_mean = Σα_i⊤_i²/Σα_i⊤_i.

Zelent B., Bialas C., Gryczynski I., Chen P., Chib R., Lewerissa K., Corradini M.G., Ludescher R.D., Vanderkooi J.M, & Matschinsky F.M. (2017). Tryptophan fluorescence yields and lifetimes as a probe of conformational changes in human glucokinase. Journal of fluorescence, 27(5), 1621-1631.

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Fluorescence Lifetime Analysis of Myc Peptide

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For biophysical validation, Myc_910–960 peptides were purchased from Thermo Fisher Scientific with 90% purity. The fluorescence lifetime of Trp in Myc peptide was measured using the DeltaFlex TCSPC system (Horiba Scientific). A quartz cuvette of 1 cm × 1 cm and 1 ml volume were used to record the spectra. The excitation monochromator's wavelength was set up at 284 nm, and the emission monochromator at 345 nm. The measurement range was set up to 200 ns with 32 nm of bandpass and peak preset of 10,000 counts. LUDOX was used to correct the instrument response factor at 284-nm wavelength. 7.5 μM Myc peptide sample with L755507 and 10074-G5 was prepared in 50 mM sodium phosphate buffer (pH 7) and incubated for 10 min at 25 °C before taking the measurement. The decay curve was fitted into a biexponential decay function using Data Analysis Software provided by Horiba Scientific.

Singh A., Kumar A., Kumar P., Nayak N., Bhardwaj T., Giri R, & Garg N. (2021). A novel inhibitor L755507 efficiently blocks c-Myc–MAX heterodimerization and induces apoptosis in cancer cells. The Journal of Biological Chemistry, 297(1), 100903.

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Peptide Fluorescence Lifetime Analysis

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Fluorescence lifetime measurements were carried out for the 5 μM peptide (pH 7.0) in increasing concentration of TFE and SDS, and 20 μM peptide (pH 7.4) in DOPC and DOPS by using TCSPC (Horiba Scientific Inc.) with pulsed LED sources. The excitation wavelength was set at 284 nm, and the emission wavelength was kept at 345 nm. A Ludox solution was used for the correct instrument response factor (IRF). Fluorescence decay data was analyzed through Data Analysis Software provided by Horiba Scientific. Decay data were fitted tri-exponentially decay function with a best fit chi-squared value approaching 1.0 to calculate the fluorescence lifetimes.

Kumar A., Kumar A., Kumar P., Garg N, & Giri R. (2021). SARS-CoV-2 NSP1 C-terminal (residues 131–180) is an intrinsically disordered region in isolation. Current Research in Virological Science, 2, 100007.

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Anisotropy Decay Measurement of Protein-Compound Interactions

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The anisotropy decay was measured using the DeltaFlex TCSPC system (Horiba Scientific). A quartz cuvette of 1 cm × 1 cm and 1 ml volume were used to record the spectra. The measurement range was set up to 200 ns with 16 nm of bandpass, peak preset of 1000 counts, and repetition rate of 1 MHz. The protein sample (20 μM Myc) with compound L755507 was incubated for half an hour at 25 °C before taking the measurement. LUDOX was used for correcting the instrument response factor at 284-nm wavelength. The anisotropy decay curve was fitted into two exponential decay function using Data Analysis Software provided by Horiba Scientific. The anisotropy decay curve can be expressed by the following equation (33 ).

Equation1.

Where, r(t) is the anisotropy at time t, r₀ is the intrinsic or initial anisotropy, A₁ and A₂ are the amplitude associated with the rotational correlation time

θ_{1}

and

θ_{2}

, respectively.

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