User:Laura Flynn/Notebook/Experimental Biological Chemistry/2011/10/05: Difference between revisions

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==Results==
==Results==
The corrected intensity of Ru(bpy)(phen) was graphed against wavelength and determined to be:
<center>[[Image:RUInt.jpg]]</center>
The integration intensity of Ru(bpy)(phen) in '''Group 3''' based on each variant concentration was determined to be:
<center>[[Image:Intensity3.jpg]]</center>
<center>[[Image:IntIntensity.jpg]]</center>
These corrected integrated intensity values show that at a concentration of about 60 μM, the peak levels out, indicating that Ru(bpy)(phen) cannot emit more light and is fully saturated.


The final concentration of MBP in each of the stock solutions was determined to be:
The final concentration of MBP in each of the stock solutions was determined to be:

Revision as of 12:33, 30 November 2011

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Objective

The purpose of today's experiment was to use standard concentrations Ru(bpy)(phen) to determine the sensitivity of a fluorimeter. This was then used to determine the concentration of a stock solution of maltose binding protein (MBP).

Procedure

  • 5 varying concentrations of MBP were prepared using TRIS as the solvent. The 5 solutions each were composed of:

So that each following stock solution had a dilution of MBP by a factor of 10.

  • These solutions were then analyzed using both UV-Vis and a fluorimeter. The intensities were taken from 310nm-500nm using an excitation value of 290nm.
  • The data from the UV-Vis was analyzed and a graph containing absorbance vs. concentration was created.
  • The absorbance of each of the solutions at 280 nm was used to calculate the concentration of MBP. This was done using the epsilon value found from [1], which was determined to be 334579.807 (1/M*cm) at 280 nm. The corresponding peak from these calculations was:
  • A graph of intensity/concentration of MBP vs. wavelength for each solution was created and used for analysis.

Results

The final concentration of MBP in each of the stock solutions was determined to be:

This was determined using the values from the UV Vis analysis. The resulting absorbance vs. wavelength graph yielded:

Each solution (A-E) was graphed as Intensity/Concentration vs. Wavelength.


The resulting graphs of both Solution A and Solution B show that the concentration of the MBP was too high to be detected on the fluorimeter.

Each of these graphs had a maximum that was determined to be:

  • The values for Stock Solutions A and B should not be factored in as their peaks were unchartable.

This graph shows the collective data of every group participating in this experiment. This graph will be updated as more groups record their information. Group 3, which is denoted by asterisks, is the group that this notebook's information is from.

Notes

  • A baseline change was preformed for the data results from Stock E. An value of 0.004 was added to each data point to prevent a negative concentration.



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