User:Eleni N. Kalivas/Notebook/CHEM-571/2013/09/10
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The objective for today was to ascertain the concentration of Adenosine Deaminase via analysis of the UV-vis spectra. Since protein concentrations are difficult to measure Bradford Assay, which is a protein binding dye, will be used. The specific dye used is called Coomassie Blue. Once the dye binds with the protein it undergoes absorption changes. We will be making a calibration curve using the Bradford Assay data for the protein.
The protocol called for 10mg of protein in 2mL of solution. Due to insufficient protein we were only about to use 3.6mg per 2mL which had a final concentration of 1,800 μg/mL. This was then used to create six standard solutions:
Absorbance Spectra of Adenosine Deaminase
The graph includes a key where starting at number 1 (our highest concentration), until 6 (which is our lowest).
Atomic Absorption Standards Prep
The following standard molarities were prepared: