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Steady-state fluorescence measurements

HK Hiyam El Kojok
ND Nada El Darra
MK Mahmoud Khalil
AC Alessandro Capo
AP Angela Pennacchio
MS Maria Staiano
AC Alessandra Camarca
SD Sabato D’Auria
AV Antonio Varriale
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Steady-state measurements were performed using a fluorescence spectrometer FP-8600 (Jasco-Japan). The polarization emission spectra of CPFX-GlnBP-CF64 were acquired setting excitation value at 650 nm and recording between 654 nm and 800 nm. In antibody binding experiments, the fluorescence spectra were carried out through incubation, overnight at 37 °C, of CPFX-GlnBP-CF647 with increasing concentrations of anti-CPFX. The antibody concentration tested was from 20 pM to 2000 pM in 0.01 mol/L PBS buffer at pH 7.4. The polarization fluorescence measurements were obtained with polarized filter, in excitation and emission, set at vertical position.

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