User:Michael S. Bible/Notebook/571/2014/09/30: Difference between revisions

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==Description==
==Description==
# CaCl<sub>2</sub> Calibration Curve
#*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)
# Cl<sup>-</sup> calibration curve
#*Prepare 25 mL 50 mM NaCl using HPLC water
#*Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM NaCl
#*Measure Cl<sup>-</sup> using ISE
#*Try measuring 50 μM and 50 mM of other salts to check for interferences & ion-pairing
#*Be sure to measure HPLC water, DI water, tap water, and your lysozyme stock solution.
# Dialysis
#*Prepare 10 mL 0.5 g/L Lysozyme
#*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


#Ca<sup>2+</sup> calibration curve
#*Prepared 25 mL 50 mM CaCl<sub>2</sub> using HPLC water
#*Using serial dilutions, prepared 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM CaCl<sub>2</sub>
#*Measured Ca<sup>2+</sup> using ISE
#*Measured 50 mM of other salts (MgCl<sub>2</sub>, CuCl<sub>2</sub>, KCl, and NaCl) added to 50 mM CaCl<sub>2</sub> to check for interferences.  Also added some HCl solution.
#*Measured HPLC water, DI water, tap water, and our lysozyme stock solution.
#Cl<sup>-</sup> calibration curve
#*Prepared 25 mL 50 mM NaCl using HPLC water
#*Using serial dilutions, prepared 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM NaCl
#*Measured Cl<sup>-</sup> using ISE
#*Measured 50 mM of other salts to check for interferences & ion-pairing
#*Also measured HPLC water, DI water, tap water, and your lysozyme stock solution.


The protocol above was adapted from [[User:Douglas_M._Fox/Notebook/AU_CHEM-571_F2011_Lab_Support/2014/09/30|Dr. Fox's Notebook]].


==Data==
==Data==

Revision as of 17:29, 1 October 2014

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Objective

Created a create a Ca2+ calibration curve and began a new dialysis

Description

  1. Ca2+ calibration curve
    • Prepared 25 mL 50 mM CaCl2 using HPLC water
    • Using serial dilutions, prepared 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM CaCl2
    • Measured Ca2+ using ISE
    • Measured 50 mM of other salts (MgCl2, CuCl2, KCl, and NaCl) added to 50 mM CaCl2 to check for interferences. Also added some HCl solution.
    • Measured HPLC water, DI water, tap water, and our lysozyme stock solution.
  2. Cl- calibration curve
    • Prepared 25 mL 50 mM NaCl using HPLC water
    • Using serial dilutions, prepared 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM NaCl
    • Measured Cl- using ISE
    • Measured 50 mM of other salts to check for interferences & ion-pairing
    • Also measured HPLC water, DI water, tap water, and your lysozyme stock solution.

The protocol above was adapted from Dr. Fox's Notebook.

Data

  • For the 0.5 g/L Lysozyme, 0.00570g Lysozyme used.
  • For NaCl serial dilution 0.0713g NaCl was placed in 25 mL of water
  • For the 0.25 mM HCl, a new solution was made containing HPLC water rather than DI water


The figure below shows the calibration curve for CaCl2. A logarithmic scale was used because the concentration values of the salt vary by an order of magnitude between each value.

In the figure below, the values of the potential were taken after the addition of ions. If the ion added first did not affect the value of the potential, the next ion was added to that same solution. As can be seen below, there is essentially no change in the potential value measured after the addition of any of the ions. This means that the presence of other ions has no effect on the potential reading of CaCl2.

Notes

This area is for any observations or conclusions that you would like to note.


Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.




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