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