Fluorescence polarization/anisotropy

Benefit description

Anisotropy measurements provide information on the size and shape of proteins or the rigidity of various molecular environments. Anisotropy measurements have been used to measure protein-protein associations or fluidity of membranes. Equipment: Modular Spectrofluorimeter Fluorolog, (Jobin Yvon - Spex). Temperature control: peltier unit.

Benefit description

Fluorescence intensity and anisotropy/polarization measurements allow the detection and quantification of molecular interactions such as those involving proteins, nucleic acids, peptides or ligands with high sensitivity and low sample consumption. Anisotropy is particularly suitable for this purpose, as it depends on molecular volume and hence, it is sensitive to changes in size that occur upon complexation. The high sensitivity of these measurements (picomolar) enables evaluation of high affinity interactions under true equilibrium conditions. By using a plate reader, measurements are fast and can be performed simultaneously on hundreds of samples of low volume (a few microliters), opening the possibility of high throughput screening applications of interest in biotechnology and biomedicine. The Facility provides acces, with the guidance and support of the staff, to a Spark® Multimode microplate reader (Tecan) equipped with polarizers for anisotropy measurements and with filters for wavelength selection [Ex: 360 (35), 485 (20), 530 (25), 560 (20), 620 (20). Em: 340 (20), 465 (35), 535 (25), 595 (35), 620 (20), 680 (30)].