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High Throughput Plate Reader Service
Synergy H4 Hybrid Multi-Mode Microplate Reader
As a result of the illumination, the sample emits light (it fluoresces) and a second optical system collects the emitted light, separates it from the excitation light (using a filter or monochromator system), and measures the signal using a light detector.
Filters/mirrors: Fluorescein 1 pM typical (0.1 fmol/well; 384-well plate)
Fluorescence Polarization can be used to measure the binding constants and kinetics of reactions that cause a change in the rotational time of the molecules. If the fluorophore is bound to a small molecule, the rate at which it tumbles can decrease significantly when it is bound tightly to a large protein. If the fluorophore is attached to the larger protein in a binding pair, the difference in polarization between bound and unbound states will be smaller
HTRF (TR-FRET) combines two techniques FRET (Fluorescence Resonance Energy Transfer) which is homogeneous and TRF (Time Resolved Fluorescence) which is low background. Advantages of TR-FRET: high sensitivity, high throughput, low false-positive, reliable results and easy protocol.
Detect the emission of cold body strong light signal by a substance not resulting from heat.
Molecules containing π-electrons or non-bonding electrons (n-electrons) can absorb the energy in the form of ultraviolet or visible light to excite these electrons to higher anti-bonding molecular orbitals. The more easily excited the electrons (i.e. lower energy gap between the HOMO and the LUMO), the longer the wavelength of light it can absorb.