Spectrophotometry - Spectrophotometry Aim and Importance:...

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Spectrophotometry Aim and Importance: There are many ways to measure the concentration of proteins but one easy way to measure small amounts of protein is colorimetric spectrophotometry. Knowing how the process of colorimetric spectrophotometry is a must for a biochemist. Background: Colorimetric spectrophotometry is done by the use of a coomassie blue hydrophobic dye called BioRad reagent, which can bind to the hydrophobic portions of the proteins. Coomassie blue has maximum absorption of light at 465nm in visible range. The maximum absorption shifts to 595nm when coomassie blue binds to protein. By generating a standard curve (concentration vs. absorbance) using bovine serum albumin (BSA) as a standard, we can calculate protein concentration by measuring the absorbance. Only a very small amount of proteins is required and it does not have to have Tyr, Try or Phe unlike other methods. However the proteins are denatured in the process. The solutions are place in the spectrophotometer, a device for measuring light
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This note was uploaded on 10/25/2011 for the course ALL 101 taught by Professor All during the Spring '11 term at Cameron University.

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Spectrophotometry - Spectrophotometry Aim and Importance:...

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