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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*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''' | *'''Ca<sup>2+</sup> calibration curve''' | ||
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[[Image:CalciumCC.png|337x196px|]]<br.> | |||
**Try measuring 50 μM and 50 mM of other salts to check for interferences (Mg<sup>2+</sup> should, Na<sup>+</sup> should not) | **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. | **Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution. | ||
{| {{table}} | {| {{table}} | ||
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|} | |} | ||
[[Image:ClCC.png]] | |||
**Try measuring 50 μM and 50 mM of other salts to check for interferences & ion-pairing | **Try measuring 50 μM and 50 mM of other salts to check for interferences & ion-pairing | ||
{| {{table}} | {| {{table}} | ||
| align="center" style="background:#f0f0f0;"|'''NaCl concentration''' | | align="center" style="background:#f0f0f0;"|'''NaCl concentration''' | ||
| align="center" style="background:#f0f0f0;"|''' | | 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)''' | | align="center" style="background:#f0f0f0;"|'''mV measurement (mV)''' | ||
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| 50mM || 50 mM||- | | 50mM || 50 mM||0||-27.0 | ||
|- | |||
| 50mM || 0||50mM||-30.8 | |||
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|} | |} | ||
**Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution. | **Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution. | ||
{| {{table}} | {| {{table}} | ||
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|} | |} | ||
===Dyalisis=== | ===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
|