User:Monika Gasiorek/Notebook/CHEM-571 2014F/2014/10/15

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October 15, 2014

Tornado Warning

Disruptive...

SDS-PAGE: Running of the Gels

The four samples prepared yesterday were heated to 40C on a heating block. Afterwards, they were transferred into a premade gel along with a premade marker/ladder. The gel was then run under the same conditions as done earlier in the semester. (See notes from September 23, 2014)

The following table depicts the placement of our samples in the 12-well gel:

Well Sample
1 Ladder
8 30:1 [Au]:[BSA] colloid
9 0.6 g/L lysozyme
10 0.12 g/L lysozyme
11 Unknown B

And the rest is history...

Primary Experiment Dialysis #2 Work-up

30:1 colloid solution vs CaCl2 using MWCO 3500

Well 1 Well 2 Well 3 Well 4 Well 5
30:1 colloid 30:1 colloid 30:1 colloid 30:1 colloid 30:1 colloid
50 mM CaCl2 5 mM CaCl2 0.5 mM CaCl2 50 μM CaCl2 5 μM CaCl2

Necessary Analyses:

  • Bradford - completed, results need to be uploaded
    • 5 colloid samples, 30:1 colloid reference, and Bradford blank
    • 20 μL sample, 200 μL 1:4 Bradford, 780 μL 50 mM Tris/50 mM NaCl

NOTE: There was a bit of a struggle getting a reliable spectrum for the blank. Was run 3 times in order to average/verify. We also made the executive decision to run samples of 50 μL from now on (diluted with 200 μL Bradford and 750 μL Tris/NaCl) in order to increase the precision/accuracy of the results.

  • Fluorescence - see 10/21
    • 5 colloid samples, 30:1 colloid reference
    • 10 μL of sample in 990 μL of DI water
  • UV-Vis - see 10/21
    • 5 colloid samples, 30:1 colloid reference
    • 10 μL of sample in 990 μL of DI water
  • CaCl2 ISE - see 10/21
    • All 10 samples

Primary Experiment Dialysis #3 Set-Up

0.6 g/L Lysozyme vs CaCl2 using MWCO 3500

Well 1 Well 2 Well 3 Well 4 Well 5
0.6 g/L lysozyme 0.6 g/L lysozyme 0.6 g/L lysozyme 0.6 g/L lysozyme 0.6 g/L lysozome
50 mM CaCl2 5 mM CaCl2 0.5 mM CaCl2 50 μM CaCl2 5 μM CaCl2