User:Alicia Rasines Mazo/Notebook/CHEM-571 Experimental Biological Chemistry/2014/10/21: Difference between revisions
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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> | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''KI concentration''' | |||
| align="center" style="background:#f0f0f0;"|'''mV measurement (mV)''' | |||
|- | |||
| 2mM || -198.3 | |||
|- | |||
| 5 mM || -217.3 | |||
|- | |||
| 10 mM || -237.0 | |||
|- | |||
| 25 mM || -247.4 | |||
|- | |||
| 50mM || -278.0 | |||
|- | |||
|} | |||
[[Image:I CC.png]]<br.> | |||
===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.> | |||
<u>Ca2+ ISE</u> | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''CaCl<sub>2</sub> concentration''' | |||
| align="center" style="background:#f0f0f0;"|'''mV measurement (mV)''' | |||
|- | |||
| 5μM (1) || 26.2 | |||
|- | |||
| Colloid (1) || 14.8 | |||
|- | |||
| 50μM || 24.5 | |||
|- | |||
| Colloid (2) || 30.8 | |||
|- | |||
| 500μM || 59.3 | |||
|- | |||
| Colloid (3) || 45.8 | |||
|- | |||
| 5mM || 70.6 | |||
|- | |||
| Colloid (4) || 66.9 | |||
|- | |||
| 50mM || 86.8 | |||
|- | |||
| Colloid (5) || 85.2 | |||
|- | |||
|} | |||
<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. | |||
<u>Fluorescence measurements</u> | |||
*There was no fluorescence since there was no free protein in solution |
Revision as of 12:54, 22 October 2014
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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
UV-Vis measurements
Fluorescence measurements
|