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12.2.1 Chromophores in proteins The electronic transitions of thepeptide bondoccur in the far UV. The intense peak at 190 nm, an ...

Molecules such as FAD (flavin adenine dinucleotide), NADH and NADþare impor- tantcoenzymesof proteins involved in electron trans ...

intensityI 0 passes through a sample with appropriate transparency and the path length (thickness)d, the intensityIdrops along t ...

Factors affecting UV/Vis absorption Biochemical samples are usually buffered aqueous solutions, which has two major advantages. ...

Alternatively, one can record the control spectrum first and use this as internal reference for the sample spectrum. The latter ...

A prism splits the incoming light into its components by refraction. Refraction occurs because radiation of different wavelength ...

detectors. At about 350 nm most instruments require a change of the light source from visible to UV light. This is achieved by m ...

by computers which allows easy processing and handling of data. From a purist’s point of view, the direct measurement of the dif ...

Difference spectra have three distinct features as compared to absolute spectra: difference spectra may contain negative absorb ...

include the binding of a monoclonal antibody to its antigen with second-order derivatives and the quantification of tryptophan a ...

In these diagrams, energy transitions are indicated by vertical lines. Not all transitions are possible; allowed transitions are ...

Since radiative energy is lost in fluorescence as compared to the absorption, the fluorescent light is always at a longer wavele ...

low concentrations. One major factor adding to the high sensitivity of fluorescence applications is the spectral selectivity. Du ...

12.3.3 Applications There are many and highly varied applications for fluorescence despite the fact that relatively few compound ...

When interpreting effects observed in fluorescence experiments, one has to consider carefully all possible molecular events. For ...

is thus a valuable tool for assessment of the degree of non-polarity. It can also be used in competition assays to monitor bindi ...

between the base pairs of double-helical DNA. Their fluorescence emission in water is very weak and increases about 30-fold upon ...

intrinsic fluorescence of the protein. While this is a very convenient titration assay when validated for an individual protein– ...

Bioluminescence Resonance Energy Transfer (BRET) Bioluminescence resonance energy transfer(BRET) uses the FRET effect with nativ ...

luciferase, which oxidises luciferin (coelenterazin) by simultaneously emitting light at lexc= 480 nm. This light directly excit ...