User:Michael S. Bible/Notebook/571/2014/10/08: Difference between revisions

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* Remove 20 μL of solution from each chamber (10 in all) and run Bradford analysis
* Remove 20 μL of solution from each chamber (10 in all) and run Bradford analysis
**Bradford reagent should be diluted 1:4 with 50mM Tris/50mM NaCl
**Bradford reagent should be diluted 1:4 with water.
**Recall, 20 μL solution + 200 μL diluted Bradford + 780 μL Tris/NaCl buffer
**Recall, 20 μL solution + 200 μL diluted Bradford + 780 μL Tris/NaCl buffer
**PS cuvettes, measuring 400 - 800 nm
**PS cuvettes, measuring 400 - 800 nm

Revision as of 14:41, 19 October 2014

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Run Bradford Analysis on Post-Dialysis Solutions

Post-dialysis solutions from all of the wells in the dialysis chamber were analyzed using UV-Vis using the following protocol:

  • Remove 20 μL of solution from each chamber (10 in all) and run Bradford analysis
    • Bradford reagent should be diluted 1:4 with water.
    • Recall, 20 μL solution + 200 μL diluted Bradford + 780 μL Tris/NaCl buffer
    • PS cuvettes, measuring 400 - 800 nm
    • Run a blank with just Bradford & buffer


Procedure taken from Dr. Fox

Description

  1. Add experimental record here. Include what, how, and why...

Data

Ca2+ ISE Data

Above are the final concentrations of Ca2+ in the post-dialysis solutions. The final two initial concentrations do not give good data as the final concentrations measured between the protein and ion side add to a greater concentration of ions than was originally present in the initial solution. These two data points should be disregarded and Lysozyme should be dialyzed against additional concentrations of Calcium ions in order to form a better curve later.


Above are the results of Bradford anlysis of the post-dialysis protein solutions. The data does not look good, and suggests that there is no protein present. The most likely reason for such data is that the wrong side of the dialysis chamber was tested using Bradford Analysis. The samples are still stored from this experiment, so it may be possible to re-run this UV-Vis experiment.

Notes

This area is for any observations or conclusions that you would like to note.


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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