User:Khyra A. Neal/Notebook/Chem 571/2014/09/30: 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"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|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]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
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| colspan="2"|
| colspan="2"|
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==Tasklist==
==September 30, 2014==
 
===Tasklist===


1. Calibration Curve for CaCl<sub>2</sub>
1. Calibration Curve for CaCl<sub>2</sub>
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* Prepare 25 mL of each using serial dilutions: 5 mM, 500 μM, 50 μM, and 5 μM CaCl<sub>2</sub> from 50 mM stock CaCl<sub>2</sub>
* Prepare 25 mL of each using serial dilutions: 5 mM, 500 μM, 50 μM, and 5 μM CaCl<sub>2</sub> from 50 mM stock CaCl<sub>2</sub>
** Dilute all solutions to 25 mL with HPLC water.
** Dilute all solutions to 25 mL with HPLC water.
*Measure Cl<sup>-</sup> using ISE
*Measure Ca<sup>2+</sup> using ISE
 
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''CaCl<sub>2</sub> concentration'''
| align="center" style="background:#f0f0f0;"|'''Potential (mV)'''
|-
| 5μM || -44.6
|-
| 50μM || 8.2
|-
| 500μM ||39.5
|-
| 5mM ||63.5
|-
| 50mM || 86.6
|-
|}
 
[[Image:CaCl2.jpg]]
 
*Measure HPLC water, DI water, tap water, and your lysozyme stock solution.
 
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Substance'''
| align="center" style="background:#f0f0f0;"|'''Potential (mV)'''
|-
| Tap water || 51.3
|-
| HPLC water || -23.0
|-
| DI water ||-51.5
|-
| Lysozyme 0.5g/L || -19.1
|-
|}
 
* Measure 50 mM and 50 mM of other salts to check for interferences
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''CaCl<sub>2</sub> concentration'''
| align="center" style="background:#f0f0f0;"|'''Cu(II)Cl<sub>2</sub> concentration'''
| align="center" style="background:#f0f0f0;"|'''KCl concentration'''
| align="center" style="background:#f0f0f0;"|'''Potential (mV)'''
|-
| 50mM || 0||50mM||86.4
|-
| 50mM || 50mM||0||80.5
|}




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*Prepare 25 mL of each using serial dilutions: 5 mM, 500 μM, 50 μM, and 5 μM NaCl from 50 mM NaCl stock solution
*Prepare 25 mL of each using serial dilutions: 5 mM, 500 μM, 50 μM, and 5 μM NaCl from 50 mM NaCl stock solution
**Dilute all solutions to 25 mL with HPLC water
**Dilute all solutions to 25 mL with HPLC water
*Measure Ca<sup>2+</sup> using ISE  
*Measure Cl<sup>-</sup> using ISE  
 
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''NaCl concentration'''
| align="center" style="background:#f0f0f0;"|'''Potential (mV)'''
|-
| 5μM || 146.5
|-
| 50μM || 129.6
|-
| 500μM ||81.3
|-
| 5mM ||27.5
|-
| 50mM || -23.9
|-
|}
 
[[Image:NaCl.jpg]]
 
*Measure HPLC water, DI water, tap water, and your lysozyme stock solution.


{| {{table}}
{| {{table}}
| align="center" style="background:#f0f0f0;"|
| align="center" style="background:#f0f0f0;"|'''Substance'''
| align="center" style="background:#f0f0f0;"|'''Conductivity'''
| align="center" style="background:#f0f0f0;"|'''Potential (mV)'''
| align="center" style="background:#f0f0f0;"|
|-
| Tap water || 65.5
|-
|-
| Concentration||NaCl||CaCl<sub>2</sub>
| HPLC water || 163.5
|-
|-
| 5 μM||146.5 mV||44.6 mV
| DI water ||158.8
|-
|-
| 50 μM||129.6 mV||8.2 mV
| Lysozyme 0.5g/L || 89.4
|-
|}
 
 
3. Try measuring 50 mM and 50 mM of other salts to check for interferences
 
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''NaCl concentration'''
| align="center" style="background:#f0f0f0;"|'''NaSO<sub>4</sub>'''
| align="center" style="background:#f0f0f0;"|'''Na<sub>2</sub>HPO<sub>4</sub><sup>2-</sup>'''
| align="center" style="background:#f0f0f0;"|'''Potential (mV)'''
|-
|-
| 500 μM||81.3 mV||39.5 mV
| 50mM || 50 mM||0||-27.0
|-
|-
| 5 mM||27.5 mV||63.5 mV
| 50mM || 0||50mM||-30.8
|-
|-
| 50 mM||23.9 mV||86.6 mV
|}
|}
===Dialysis===
* Prepare 10 mL 0.5 g/L Lysozyme
** 0.005 g Lysozyme in 25 mL HPLC water using volumetric flask
* Prepare 0.25 mM HCl from 2.5 mM HCl stock solution
** 10x dilution from stock
* Cut a piece of 3500 MWCO flat dialysis tubing, wet it, and insert it in the 5-well dialysis chamber and clamp the dialysis chamber shut.
* Fill one side of each well with 1 mL of  0.5 g/L Lysozyme solution
** Measure out 1 mL with transfer pipette then add 1mL using Pasteur pipette to the wells
* Fill the other side of the wells with the following solutions:
** HPLC water, 0.25 mM HCl, 50 mM CaCl<sub>2</sub>, 500 μM CaCl<sub>2</sub>, and 50 mM NaCl
*Insert screws to prevent leaving/evaporation
*Place on low speed shaker overnight.


<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

Latest revision as of 00:21, 27 September 2017

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September 30, 2014

Tasklist

1. Calibration Curve for CaCl2

  • Prepare 25 mL 50 mM CaCl2 using HPLC water
    • 0.13875 g CaCl2 in 25 mL HPLC water
  • Prepare 25 mL of each using serial dilutions: 5 mM, 500 μM, 50 μM, and 5 μM CaCl2 from 50 mM stock CaCl2
    • Dilute all solutions to 25 mL with HPLC water.
  • Measure Ca2+ using ISE
CaCl2 concentration Potential (mV)
5μM -44.6
50μM 8.2
500μM 39.5
5mM 63.5
50mM 86.6

  • Measure HPLC water, DI water, tap water, and your lysozyme stock solution.
Substance Potential (mV)
Tap water 51.3
HPLC water -23.0
DI water -51.5
Lysozyme 0.5g/L -19.1
  • Measure 50 mM and 50 mM of other salts to check for interferences
CaCl2 concentration Cu(II)Cl2 concentration KCl concentration Potential (mV)
50mM 0 50mM 86.4
50mM 50mM 0 80.5


2. Calibration Curve for NaCl

  • Prepare 25 mL of 50 mM NaCl using HPLC water
    • 0.07305 g NaCl in 25 mL HPLC water
  • Prepare 25 mL of each using serial dilutions: 5 mM, 500 μM, 50 μM, and 5 μM NaCl from 50 mM NaCl stock solution
    • Dilute all solutions to 25 mL with HPLC water
  • Measure Cl- using ISE
NaCl concentration Potential (mV)
5μM 146.5
50μM 129.6
500μM 81.3
5mM 27.5
50mM -23.9

  • Measure HPLC water, DI water, tap water, and your lysozyme stock solution.
Substance Potential (mV)
Tap water 65.5
HPLC water 163.5
DI water 158.8
Lysozyme 0.5g/L 89.4


3. Try measuring 50 mM and 50 mM of other salts to check for interferences

NaCl concentration NaSO4 Na2HPO42- Potential (mV)
50mM 50 mM 0 -27.0
50mM 0 50mM -30.8


Dialysis

  • Prepare 10 mL 0.5 g/L Lysozyme
    • 0.005 g Lysozyme in 25 mL HPLC water using volumetric flask
  • Prepare 0.25 mM HCl from 2.5 mM HCl stock solution
    • 10x dilution from stock
  • Cut a piece of 3500 MWCO flat dialysis tubing, wet it, and insert it in the 5-well dialysis chamber and clamp the dialysis chamber shut.
  • Fill one side of each well with 1 mL of 0.5 g/L Lysozyme solution
    • Measure out 1 mL with transfer pipette then add 1mL using Pasteur pipette to the wells
  • Fill the other side of the wells with the following solutions:
    • HPLC water, 0.25 mM HCl, 50 mM CaCl2, 500 μM CaCl2, and 50 mM NaCl
  • Insert screws to prevent leaving/evaporation
  • Place on low speed shaker overnight.



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