1601 pc spectrophotometer

¹H Nuclear magnetic resonance (NMR) spectra were recorded on Bruker Avance AV300 (300 MHz) NMR instrument using CDCl₃ as the solvent. The IR spectra were recorded in the range of 4000–400 cm⁻¹ using a Bruker ALPHA FT-IR Spectrophotometer with a resolution of 4.0 cm⁻¹. Particle morphology was examined using scanning electron microscopy (JEOL JSM-6701F). The UV/Vis absorption studies were performed on a Shimadzu-1601 PC spectrophotometer. The steady state fluorescence studies were carried out on an Agilent Cary Eclipse Fluorescence Spectrophotometer.
For quantum yield measurement, following Equation (4) was used to measure relative quantum yield of particles. $$\mathit{\Phi }={\mathit{\Phi }}_R\mathrm{x}\frac{I}{I_R}x\frac{A_R}{A}\mathrm{x}\frac{{\eta }^2}{{\eta }_R^2}$$ where, Φ_R, I_R, A_R and η_R refers to quantum yield, integrated area under emission spectra, absorbance and refractive index of reference, Rhodamine B (ΦR  =  0.7 and η = 1.36) in ethanol^{45 (link)} solvent, I, A and η refers to integrated area under emission spectra, absorbance and refractive index of sample in THF solvent.

The CO-reduction assay of CYP125A13 was conducted according to a defined protocol [30 (link)]. The protein was diluted in potassium phosphate buffer (50 mM) containing 10% glycerol. A small amount of sodium dithionite was added and the solution was divided into two cuvettes. One cuvette was used as a reference while the other was saturated with 60–70 CO bubbles at a rate of one bubble per second. The solution was then scanned between 400 and 500 nm at room temperature using a Shimadzu 1601PC spectrophotometer. The CYP concentration was calculated by the difference in the absorbance at 450 nm and 490 nm using an extinction coefficient of 91 mm^-1cm^-1.