Probing bis-ANS Binding Sites of Different Affinity on Aggregated IgG by Steady-State Fluorescence, Time-Resolved Fluorescence and Isothermal Titration Calorimetry

2011 ◽  
Vol 100 (4) ◽  
pp. 1294-1305 ◽  
Author(s):  
Andrea Hawe ◽  
Theo Rispens ◽  
James N. Herron ◽  
Wim Jiskoot
2013 ◽  
Vol 85 (3) ◽  
pp. 589-608 ◽  
Author(s):  
Marcel Ameloot ◽  
Martin vandeVen ◽  
A. Ulises Acuña ◽  
Bernard Valeur

After recalling the basic relations relevant to both steady-state and time-resolved fluorescence polarization, it is shown how the values of steady-state polarized intensities recorded experimentally usually need to be corrected for systematic effects and errors, caused by instrumentation and sample properties. A list of selected reference values of steady-state fluorescence anisotropy and polarization is given. Attention is also paid to analysis of time-resolved fluorescence anisotropy data obtained by pulse fluorometry or phase and modulation fluorometry techniques. Recommendations for checking the accuracy of measurements are provided together with a list of selected time-resolved fluorescence anisotropy data as reported in the literature.


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