I have a cysteine-rich domain in my recombinant protein. results of SDS-page in reducing and non-reducing condition improved the presence of oligomers caused by disulfide bond between monomer protein in sample. the tryptophan emission spectra of my protein shows two peek in 330 and 380nm. is that true to attribute second peek to the oligomers?
The 330 nm peak is consistent with Trp in a hydrophobic environment, but even in an aqueous environment, the peak emission of Trp is not going to exceed 350 nm, so I don't think the 380 nm peak is from Trp. What happens to the 380 nm peak when you add reducing agent?
actually I have 10mM BME in protein solution always,
I titrated the sample with both ME and DTT, and saw the blue shift from 380nm to 370 nm, but the intensity and the form of spectra was changed spatially in high concentration of ME.
Is it a peak at 380nm or just a shoulder? Can you post the spectrum? Also what instrument are you using to obtain the emission spectrum?
I think this is more bigger to consider a shoulder. but I'm not sure. i will send you .Cary Eclipse fluorimeter.
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