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|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab</span> | |style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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== | ==Analysis of Dialysis Begun Yesterday== | ||
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 | |||
**Don't forget to run a blank with just Bradford & buffer | |||
**Don't forget to run your undialyzed Lysozyme stock | |||
Procedure taken from [[User:Douglas_M._Fox/Notebook/AU_CHEM-571_F2011_Lab_Support/2014/10/01|Dr. Fox]] | |||
==Data== | ==Data== | ||
[[Image:Bradford_Analysis_of_All_Post_Dialysis_Solutions.png]] | |||
This | |||
Figure 1 shows the corrected UV-Vis Bradford Spectra for all Post-Dialysis Solutions. | |||
[[Image:Bradford_analysis_of_post_dialysis_solutions_from_non_protein_side_of_dialysis_chamber.png]] | |||
Figure 2 shows the corrected UV-Vis Bradford Spectra for the post-dialysis solutions from the non-protein side of the dialysis chamber. From the negative absorbance values, it is clear that there is no protein present. This is expected since the dialysis tubing used had a molecular weight cut-off of 3500 g/mol. | |||
[[Image:Bradford_Analysis_of_post_dialysis_solutions_from_protein_side_of_dialysis_chamber.png]] | |||
Figure 3 shows the corrected UV-Vis Bradford Spectra for the post-dialysis solutions from the protein side of the dialysis chamber. It is clear from the spectra that there is still a lot of protein in each of the chambers, and the post-dialysis concentration of Lysozyme in solution almost perfectly matches the concentration of Lysozyme in the stock solution. | |||
[[Image:Lysozyme_Concentrations_via_Bradford_Post_dialysis.png]] | |||
Figure 4 shows the calculations done to determine the concentration of Lysozyme in each solution. From these calculations, it would seem that the actual concentration of stock Lysozyme solution is .28 g/L rather than .5 g/L as believed. This is because the stock solution is diluted by a factor of 50 when it is mixed with Bradford reagent and buffer. | |||
[[Category:Course]] | [[Category:Course]] | ||
Latest revision as of 00:26, 27 September 2017
 Biomaterials Design Lab
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Analysis of Dialysis Begun YesterdayPost-dialysis solutions from all of the wells in the dialysis chamber were analyzed using UV-Vis using the following protocol:
Procedure taken from Dr. Fox Data
Figure 1 shows the corrected UV-Vis Bradford Spectra for all Post-Dialysis Solutions.
Figure 2 shows the corrected UV-Vis Bradford Spectra for the post-dialysis solutions from the non-protein side of the dialysis chamber. From the negative absorbance values, it is clear that there is no protein present. This is expected since the dialysis tubing used had a molecular weight cut-off of 3500 g/mol.
Figure 3 shows the corrected UV-Vis Bradford Spectra for the post-dialysis solutions from the protein side of the dialysis chamber. It is clear from the spectra that there is still a lot of protein in each of the chambers, and the post-dialysis concentration of Lysozyme in solution almost perfectly matches the concentration of Lysozyme in the stock solution.
Figure 4 shows the calculations done to determine the concentration of Lysozyme in each solution. From these calculations, it would seem that the actual concentration of stock Lysozyme solution is .28 g/L rather than .5 g/L as believed. This is because the stock solution is diluted by a factor of 50 when it is mixed with Bradford reagent and buffer.
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