User:Klare Lazor/Notebook/Chem-496-001/2011/09/14: Difference between revisions

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==Data==
==Data==
[[Image:Asept14.png]]
[[Image:Data_sept_14.png]]
 
Image 1: Is a linear fit of concentration vs. absorbance for the wavelength of 495nm for the corrected values without the bradford reagent.
 
 
 
[[Image:Bsept14.png]]
 
Image 2: Absorbance of just the protein blank in water no bradford reagent. These values will be able to determine the concentration of the unknown protein.
 
 
 
 
 


<b>What concentration do you calculate for your unknown? What does the absorbance of just the protein in water (no Bradford reagent) look like? What is the ε value for the protein at different wavelengths?</b> [[User:Matt Hartings|Matt Hartings]] 21:48, 20 September 2011 (EDT)
<b>What concentration do you calculate for your unknown? What does the absorbance of just the protein in water (no Bradford reagent) look like? What is the ε value for the protein at different wavelengths?</b> [[User:Matt Hartings|Matt Hartings]] 21:48, 20 September 2011 (EDT)

Revision as of 10:51, 28 September 2011

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Objective

To determine the concentration of an unknown protein, a Bradford Assay was used.

Description

To determine the concentration of an unknown protein, a Bradford Assay was performed. Materials needed included Bradford Reagent, bovine serum albumen (BSA), and protein (MBP). Standard solutions of BSA (1mL each) in water of 10, 8, 6, 4, 2, and 1ug/mL were prepared. First, 3mL of a 10ug/mL stock solution was created. From this stock solution the standards, as well as the sample of unknown protein, were made. The sample of the unknown protein should be 1000 times too concentrated. Furthermore, a blank for the Bradford Reagent and the protein were prepared. A UV-visible spectra was taken of each solution with a wavelength range of 200-800nm.

Data

What concentration do you calculate for your unknown? What does the absorbance of just the protein in water (no Bradford reagent) look like? What is the ε value for the protein at different wavelengths? Matt Hartings 21:48, 20 September 2011 (EDT)

Notes

Observations:

  • It was observed that the color of the Bradford Reagent upon addition to the protein was blue, and slowly, turned back to red when the concentration of the protein decreased.


Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.


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