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FTIR spectroscopy

MK Maithili Krishnan-Schmieden
PK Patrick E. Konold
JK John T. M. Kennis
AP Anjali Pandit
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Infrared spectra were recorded using an FTIR spectrometer (IFS 66 s Bruker) equipped with a nitrogen-cooled photovoltaic Mercury Cadmium Telluride (MCT) detector (20 MHz, KV100, Kolmar Technologies) described earlier55–57. The samples were contained between CaF2 windows separated with a 20 µm Teflon spacer and the concentration was tuned for OD ~0.8 absorption at 1650 cm−1, at a protein concentration of approximately 1 mM, with a volume of 100 μl. The measurements were carried out at room temperature and spectral resolution of the instrument was 3 cm−1. The samples were solubilized in D2O detergent buffer at either pD 5.0 or pD 7.5. Measurements with the pD 5.0 and 7.5 samples were collected consecutively and difference FTIR spectra were produced by scaling according to the integrated linear absorption of the amide I band (1610–1690 cm−1).

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