User:Alicia Rasines Mazo/Notebook/CHEM-571 Experimental Biological Chemistry/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"| | |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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==Tasks for September 30== | ==Tasks for September 30== | ||
*To run dyalisis | *To run dyalisis | ||
*To prepare calibration curves Ca<sup>2+</sup> and Cl<sup>-</sup> | *To prepare calibration curves Ca<sup>2+</sup> and Cl<sup>-</sup> | ||
=== | ===Determining calibration curves for Ca<sup>2+</sup> and Cl<sup>-</sup>=== | ||
Procedure followed as described by [http://openwetware.org/wiki/User:Douglas_M._Fox/Notebook/AU_CHEM-571_F2011_Lab_Support/2014/09/30 Dr. Fox] | Procedure followed as described by [http://openwetware.org/wiki/User:Douglas_M._Fox/Notebook/AU_CHEM-571_F2011_Lab_Support/2014/09/30 Dr. Fox] | ||
*'''Ca<sup>2+</sup> calibration curve''' | |||
**Prepare 25 mL 50 mM CaCl<sub>2</sub> using HPLC water | |||
***Added 0.13876g CaCl<sub>2</sub> to a 25 mL volumetric flask and made up to the mark with HPLC | |||
**Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM CaCl<sub>2</sub> | |||
**Measure Ca<sup>2+</sup> using ISE | |||
# | {| {{table}} | ||
| align="center" style="background:#f0f0f0;"|'''CaCl<sub>2</sub> concentration''' | |||
| align="center" style="background:#f0f0f0;"|'''mV measurement (mV)''' | |||
|- | |||
| 5μM || -44.6 | |||
|- | |||
| 50μM || 8.2 | |||
|- | |||
| 500μM ||39.5 | |||
|- | |||
| 5mM ||63.5 | |||
|- | |||
| 50mM || 86.6 | |||
|- | |||
|} | |||
# | [[Image:CalciumCC.png|337x196px|]]<br.> | ||
#*Prepare 10 mL 0.5 g/L Lysozyme | **Try measuring 50 μM and 50 mM of other salts to check for interferences (Mg<sup>2+</sup> should, Na<sup>+</sup> should not) | ||
# | {| {{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;"|'''mV measurement (mV)''' | ||
# | |- | ||
# | | 50mM || 0||50mM||86.4 | ||
# | |- | ||
# | | 50mM || 50mM||0||80.5 | ||
# | |} | ||
# | '''Note''': There may be minimal interference by Cl<sup>-</sup> | ||
# | **Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution. | ||
# | {| {{table}} | ||
# | | align="center" style="background:#f0f0f0;"|'''Substance''' | ||
# | | align="center" style="background:#f0f0f0;"|'''mV measurement (mV)''' | ||
|- | |||
| Tap water || 51.3 | |||
|- | |||
| HPLC water || -23.0 | |||
|- | |||
| DI water ||-51.5 | |||
|- | |||
| Lysozyme 0.5g/L || -19.1 | |||
|- | |||
|} | |||
*'''Cl<sup>-</sup> calibration curve''' | |||
**Prepare 25 mL 50 mM NaCl using HPLC water | |||
***Added 0.07091 g NaCl to a 25 mL volumetric flask and made up to the mark with HPLC | |||
**Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM NaCl | |||
**Measure Cl<sup>-</sup> using ISE | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''NaCl concentration''' | |||
| align="center" style="background:#f0f0f0;"|'''mV measurement (mV)''' | |||
|- | |||
| 5μM || 146.5 | |||
|- | |||
| 50μM || 129.6 | |||
|- | |||
| 500μM ||81.3 | |||
|- | |||
| 5mM ||27.5 | |||
|- | |||
| 50mM || -23.9 | |||
|- | |||
|} | |||
[[Image:ClCC.png]] | |||
**Try measuring 50 μM and 50 mM of other salts to check for interferences & ion-pairing | |||
{| {{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;"|'''mV measurement (mV)''' | |||
|- | |||
| 50mM || 50 mM||0||-27.0 | |||
|- | |||
| 50mM || 0||50mM||-30.8 | |||
|- | |||
|} | |||
**Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution. | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Substance''' | |||
| align="center" style="background:#f0f0f0;"|'''mV measurement (mV)''' | |||
|- | |||
| Tap water || 65.5 | |||
|- | |||
| HPLC water || 163.5 | |||
|- | |||
| DI water ||158.8 | |||
|- | |||
| Lysozyme 0.5g/L || 89.4 | |||
|- | |||
|} | |||
===Dyalisis=== | |||
#Prepare 10 mL 0.5 g/L Lysozyme | |||
#*Added 0.0050 g to 10mL volumetric flask and made up to mark with HPLC | |||
#Prepare 2 mL 50 mM CaCl<sub>2</sub> (if you read ahead, you can use your CaCl<sub>2</sub> calibration stock) | |||
#Prepare 2 mL 500 μM CaCl<sub>2</sub> (if you read ahead, you can use a CaCl<sub>2</sub> calibration dilution) | |||
#Prepare 2 mL 50 mM NaCl (if you read ahead, you can use your NaCl calibration stock) | |||
#Prepare 1 mL 0.25 mM HCl using your 2.5 mM stored stock | |||
#Cut a piece of 3500 MWCO flat dialysis tubing, wet it, and insert it in the 5-well dialysis chamber | |||
#Clamp the dialysis chamber shut | |||
#Fill one side of each well with 1 mL of your 0.5 g/L Lysozyme solution | |||
#*Transfer 1 mL to a plastic test tube using a pippetter, then | |||
#*Carefully fill each well with with a glass Pasteur pipette | |||
#*You may need to tip the chamber on an angle to reduce water surface tension | |||
#Fill the other side of the well with one of the following solutions | |||
#*HPLC water, 0.25 mM HCl, 50 mM CaCl<sub>2</sub>, 500 μM CaCl<sub>2</sub>, or 50 mM NaCl | |||
#*Each well has a different solution, which will be tested tomorrow | |||
#Insert top screws to prevent leaking/evaporation | |||
<u>pH measurements</u> | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Substance''' | |||
| align="center" style="background:#f0f0f0;"|'''pH measurement''' | |||
|- | |||
| 0.25 mM HCl (diluted 10x) (2) || 7.857 | |||
|- | |||
| Lysozyme (diluted 10x) (2) || 7.426 | |||
|- | |||
|} |
Latest revision as of 00:21, 27 September 2017
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Tasks for September 30
Determining calibration curves for Ca2+ and Cl-Procedure followed as described by Dr. Fox
Note: There may be minimal interference by Cl-
Dyalisis
pH measurements
|