My protein shows good signal in far UV and fluorescence spectra but in case of near UV there is very less/ no signal even at high concentration of protein.
what will be the possible explanation for this?
any supporting paper to explain this
Mr. Bansal,
Could you provide more detail about the Fs spectra?
Excitation wavelength was 290nm
Emission wavelength was 340nm,
Scan range 300-- 400nm,
Showing nice peak
Aromatic amino acids (Phe, Tyr, Trp), and disulfide bonds are responsible for the UV absorbance of proteins. Bound ligands and prosthetic groups, if present, will also contribute to the absorbance and, in some cases, fluorescence. Tryptophan is mainly responsible for the fluorescence of proteins if there are no fluorescent ligands or prosthetic groups. Trp absorbs maximally at about 280 nm and emits between 330 and 355 nm, depending upon the hydrophobicity of the environment. It sounds like you are seeing Trp fluorescence. However, make sure you subtract the background caused by Raman scatter from water by taking a spectrum of the buffer under the same conditions.
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