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Objective
Created a create a Ca2+ calibration curve and began a new dialysis
Description
- CaCl2 Calibration Curve
- Prepare 2 mL 50 mM CaCl2 (if you read ahead, you can use your CaCl2 calibration stock)
- Prepare 2 mL 500 μM CaCl2 (if you read ahead, you can use a CaCl2 calibration dilution)
- Prepare 2 mL 50 mM NaCl (if you read ahead, you can use your NaCl calibration stock)
- Cl- calibration curve
- Prepare 25 mL 50 mM NaCl using HPLC water
- We calculated that 0.07305g NaCl was necessary
- Using serial dilutions, prepare 25 mL each of 5 mM, 500 μM, 50 μM, & 5 μM NaCl
- Measure Cl- 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 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
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
| NaCl Calibration Curve
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Conc. (mM)
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mV
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50 |
-24
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5 |
28.5
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0.5 |
88.2
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0.05 |
128.8
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0.005 |
152.8
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 Polyvinyl Alcohol Films with Absorbed Malachite Green Concentrations; 0.2, 0.5, 1.1, 1.4, 1.7, and 2.0 ppm
Notes
- For the 0.25 mM HCl, a new solution was made containing HPLC water rather than DI water
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Biomaterials Design Lab
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