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An excited fluorescent molecule such as Green fluorescent probe (GPF) or Yellow fluorescent probe (YFP) as a doner-acceptor pair which is dispose off the energy from the absorbed photon by non radio-active fluorescence resonance energy transfer (FRET), in which the energy of the excited molecule directly passes to a nearby molecule without emission of a photon. The emitted photon has a longer wavelength then both the original exciting light. The efficiency of FRET is inversely proportional to the sixth power of the distance between the donor and acceptor. Fluorescent probes are frequently used to detect rapid biochemical changes in a single living cell and also used to measure cAMP in living cell. These molucules are called fluorophore and this is the special quality due to tihs gies molucular information unique to tha revealed by solvent-dependent process.

FRET analysis having many advantages over any other techniques as for example isothermal titration calorimetry in which concentrations are determined accurately by absorbance, having highly sensitive binding signals enable the analysis in minimum amount.

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The various techniques through which it is achieved are discussed in brief as follows:

  1. Spectroscopy
  2. Mass Spectrometry
  3. Chromatography
  4. Electrophoresis
  5. Isoelectric Focussing
  6. Microscopy
  7. Scatchard Analysis
  8. FRET
  9. Blotting

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