Molecular Fluorescence Analysis of Proteins Lab Report

Molecular Fluorescence Analysis of Proteins - Lab announce ExampleAs the electron returns from the lowest triplet state to the singlet ground state, a photon is released. The risque sensitivity and specificity of fluorescence techniques facilitates the study of individual protein molecules.Therefore, this try sought to determine the source of fluorescence in serum albumen by first establishing the fluorescence of two key amino acids that make up serum albumin. Such a study would enable a detailed study of serum albumin as a protein. The experiment also aimed at establishing the concentration of riboflavin development the absorption of known concentrations of riboflavin.Solutions of 10 M tyrosine and tryptophan were made to volumes of 100L each by first preparing 100 mL of 1mM solutions and diluting them to 10 M. The applications bundle of the fluorescent instrument was loaded after which the condition parameters were set. An emission scan range of between 290 and 380 nm was s et alongside slits of 5nm. The emission wavelength was set to 280 nm, and the emission tab was clicked to obtain the emission spectrum of the buffer zone solution. The spectrum was rescue after which the same procedure was repeated for tryptophan. Thereafter, an excitation spectrum for tryptophan was prepared by choosing the optimal emission wavelength according to the results of the emission spectrum. A tyrosine spectrum was obtained using similar conditions as those of the buffer. The tyrosine band was then obtained by subtracting the buffer spectrum from the tyrosine spectrum. All these steps were repeated for serum albumin making sure to save all the spectra.1.0 mL of 10 ppm riboflavin solution was measured into a 100 mL volumetric flask and thin out using the buffer to make a 100 ppb stock solution. Standard solutions were then prepared using clean volumetric pipettes rinsed with HPLC water. The final concentrations of the standard solutions were