User:Beatriz Gimenez De C./Notebook/571/2014/09/30: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
No edit summary
Line 39: Line 39:
** Measure HPLC water, DI water, tap water, and your lysozyme stock solution.
** Measure HPLC water, DI water, tap water, and your lysozyme stock solution.


==Results==


<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''CaCl<sub>2</sub> Concentration (μM)'''
| align="center" style="background:#f0f0f0;"|'''mV measurement (mV)'''
|-
| 5||-44.6
|-
| 50||8.2
|-
| 500||39.5
|-
| 5000||63.5
|-
| 50000||86.6
|}
|}
__NOTOC__
__NOTOC__

Revision as of 12:08, 30 September 2014

Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Tasks

  • Make a Dialysis of lysozyme in HPLC
    • Prepare 10 mL 0.5 g/L Lysozyme (added 0.0050 g to 10 mL volumetric flask filling the rest with HPLC)
    • Prepare 2 mL 50 mM CaCl2
    • Prepare 2 mL 500 μM CaCl2
    • Prepare 2 mL 50 mM NaCl
    • Prepared 50 mL 0.25 mM HCl with HPLC
    • 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
      • 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 CaCl2, 500 μM CaCl2, or 50 mM NaCl
      • Each well has a different solution, which will be tested tomorrow
    • Insert top screws to prevent leaking/evaporation.
  • Make Ca2+ calibration curve
    • Prepare 25 mL 50 mM CaCl2 using HPLC water (0.13876 g CaCl2 in 10 mL volumetric flask with HPLC)
    • Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM CaCl2
    • Measure Ca2+ using ISE
    • Try measuring 50 μM and 50 mM of other salts to check for interferences (Mg2+ should, Na+ should not)
    • Measure HPLC water, DI water, tap water, and your lysozyme stock solution.
  • Make Cl- calibration curve
    • Prepare 25 mL 50 mM NaCl using HPLC water (0.0730 g in 10 mL volumetric flask with HPLC)
    • Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM NaCl
    • Measure Cl- using ISE
    • Measure 50 μM and 50 mM of other salts to check for interferences & ion-pairing
    • Measure HPLC water, DI water, tap water, and your lysozyme stock solution.

Results

CaCl2 Concentration (μM) mV measurement (mV)
5 -44.6
50 8.2
500 39.5
5000 63.5
50000 86.6
Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Beatriz_Gimenez_De_C./Notebook/571/2014/09/30&oldid=824911"