Eclipse cary varian uv vis spectrofluorometer

Fluorescence measurements were carried out using Eclipse Cary Varian UV-Vis spectrofluorometer (Varian, Inc. Hansen Way, Palo Alto, CA, USA) attached to a Peltier temperature controller. An excitation wavelength of 280 nm with an excitation slit of 5 nm and an emission slit of 5 nm was used and the fluorescence was recorded from 300 nm to 400 nm. Unfolding pattern and stability of protein was determined using 0–6 M of guanidine HCl (GdHCl) as described by Bansal et al., [17] . The fluorescence intensity was plotted as the function of wavelength using F₄₀₀ nm as the baseline.

The Eclipse Cary Varian UV-Vis spectrofluorometer (Varian Inc., Palo Alto, California, USA) was used to measure fluorescence and was connected to a Peltier temperature controller. To observe intrinsic tryptophan fluorescence, the protein was excited at 292 nm, and the emission of fluorescence was tracked between 300 and 400 nm (Adman and Jensen, 1981 (link)). A. lakoocha peroxidase activity was observed under different pH and chemical denaturant conditions utilizing hydrogen peroxide as a hydrogen acceptor and other substances like guaiacol as a hydrogen donor. 1 mM of peroxidase was introduced to a 50 mM acetate buffer (substrate II) containing guaiacol and 0.2 mM hydrogen peroxide (substrate I). At 470 nm, the guaiacol absorbance change rate was calculated.