I want to study the structure of my protein using intrinsic fluorescence spectroscopy. some times I see the emission peak at 380nm?
is my protein aggregated? or there is another reason for this problem?
I work on HIV-1 Tat protein that is considered as an IDP.
Does the absorption and excitation spectra (monitored at, say 400 nm) matches?
I don't know. I mean I didn't check the absorption. I just excited sample at 280&295nm and recorded the emission spectra in 300-450nm
You should do that first. In fluorescence spectroscopy it is essential that the absorption and excitation spectra matches. Mismatch of the two spectra means there are more than one molecule is present in the ground state. Most of the time the mismatch is found to be due to contamination by fluorescent molecule.
how did you analysed fluorescence spectra, without absorption spectra, tell me your exact absorption value
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