Colorimetric Methods For Quantification of Proteins

Colorimetric Methods For Quantification of Proteins - 3 Mix...

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Ashay Mehta Chemistry 124 Honors March 24, 2009 Colorimetric Method for Quantification of Proteins Introduction In this experiment, we used the Bradford method. This method works by protein which binds to the dye resulting of an alter in the absorption maximum of the dye from 465 to 595 nm in acidic solutions. This special dye interacts electro statically with amino and carboxyl groups and Van der Waals forces. Materials 6 small test tubes Dye reagent Protein solutions Spectrometer Procedure 1. See how much solution is needed to create a .25 M, .5 M, 1M, and 1.25 M concentration of the solution. 2. Create the all solutions with one blank and one composed fully of the sample.
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Unformatted text preview: 3. Mix with dye and let it sit for about five minutes. 4. Measure the absorbance of each of the solutions. 5. Create a graph of the absorbance of all the solutions. 6. Then calculate the concentrations of the unknown sample solution with the best fit line. Results Concentration (M) Absorbance (nm) Blank, 0 .25 .361 .5 .550 1 .830 1.25 .985 Sample y = 0.7357x + .1038 .714 = 0.7357x + .1038 x = 0.829 .714 (See Graph) Conclusion Using the best fit line we were able to find the concentration of the protein in the unknown sample. The graph relates the absorbance and concentrations. We learned how to effectively use the Bradford method....
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This note was uploaded on 06/02/2010 for the course MGMT 121 taught by Professor Morozova during the Spring '10 term at NJIT.

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Colorimetric Methods For Quantification of Proteins - 3 Mix...

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