User:Alicia Rasines Mazo/Notebook/CHEM-571 Experimental Biological Chemistry/2014/10/21: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</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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***The Ag/AgI wire can be connected directly to a multimeter using alligator clips | ***The Ag/AgI wire can be connected directly to a multimeter using alligator clips | ||
***You will continue to connect your Ag wire to the red lead on the multimeter | ***You will continue to connect your Ag wire to the red lead on the multimeter | ||
<u>I ISE</u> | <u>I- ISE</u> | ||
{| {{table}} | {| {{table}} | ||
| align="center" style="background:#f0f0f0;"|'''KI concentration''' | | align="center" style="background:#f0f0f0;"|'''KI concentration''' | ||
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[[Image:I CC.png]]<br.> | |||
===30:1 colloid solution vs CaCl2 (MWCO 3500) Dialysis Data=== | ===30:1 colloid solution vs CaCl2 (MWCO 3500) Dialysis Data=== | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
<u>Bradford Analysis</u> | |||
*Only running bradford analysis of protein-containing solutions, that is Colloid 1, 2, 3, 4 and 5, since the protein cannot diffuse through 3500 MWCO. | |||
* Remove 100 μL of solution from each chamber (7 in all) and run Bradford analysis | |||
**Bradford reagent should be diluted 1:3 with 50mM Tris/50mM NaCl | |||
**Recall, 100 μL solution + 200 μL diluted Bradford + 700 μL Tris/NaCl buffer | |||
**PS cuvettes, measuring 400 - 800 nm | |||
[[Image:Bradford Colloid 221014.png]]<br.> | |||
'''Note''': The colloid solution at 50 mM CaCl<sub>2</sub> was clear at the top, and precipitated colloid was sank in the bottom of the well. Because only the clear solution was extracted from the well, the protein detected using Bradford analysis corresponds to free lysozyme protein in solution. <br.> | |||
<u>Ca2+ ISE</u> | <u>Ca2+ ISE</u> | ||
{| {{table}} | {| {{table}} | ||
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<u>UV-Vis measurements</u> | <u>UV-Vis measurements</u> | ||
* No peak at 280 nm for the dialysed colloid solutions because lysozyme contains tryptophan, tyrosine, phenylalanine which are reducing agents for gold. No free protein in solution was found. | * No peak at 280 nm for the dialysed colloid solutions because lysozyme contains tryptophan, tyrosine, phenylalanine which are reducing agents for gold. No free protein in solution was found. | ||
<u>Fluorescence measurements</u> | |||
*There was no fluorescence since there was no free protein in solution |
Latest revision as of 00:27, 27 September 2017
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Tasks for Oct. 21
I- ISE preparationProdecure as describer by Dr. Fox on Oct. 21
I- ISE
30:1 colloid solution vs CaCl2 (MWCO 3500) Dialysis DataBradford Analysis
<br.> Note: The colloid solution at 50 mM CaCl2 was clear at the top, and precipitated colloid was sank in the bottom of the well. Because only the clear solution was extracted from the well, the protein detected using Bradford analysis corresponds to free lysozyme protein in solution. <br.> Ca2+ ISE
UV-Vis measurements
Fluorescence measurements
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