I mixed made solutions of PBS and Rhodamine with varying Rhodamine concentrations and ran them through the spectrometer. Below is the concentration vs absorption curve I made from it. I blanked the spec with PBS instead of ddH2O since the solution is in PBS. The slope is almost double for Rhodamine in ddH2O compared to…
Art is going to make a second stir cell this week. I asked him to replicate it as closely as possible, however I’m not sure how well that’s going to work. The one we currently have is made out of Ertalyt, a material similar to teflon. Paul gave us polyethylene which is much softer than…
I ran another diffusion cell last week using Human Albumin and Cytochrome C. I increased the volume of PBS because the last time I ran the gel for this mixture, the band for cytochrome c was very prominent. Out of the three chambers of the diffusion cell that I ran, only one of them actually…
I re-ran the rhodamine dilutions narrowing in on the molarities within the range readable by the spectrometer. My start sample was .5mM of Rhodamine. The wavelength I set the spectrometer at was 530nm with 3 trials of each dilution. I started with the least concentrated dilution and cleaned the cuvette between every trial so as…
I fixed the graph for the serial dilution of Rhodamine that I did last week and removed all absorbency values that were greater than 1.08 AU. The graph looks much more linear now with an R^2 value of .9517
I ran a series of serial dilutions with the rhodamine, to find the absorbency using the spectrometer. Below is a graph of the concentration of rhodamine versus the absorbency. I removed the concentrations which had absorbances greater than 1.7 AU because those absorbencies created a greater deviation. The lower concentrations show a more linear distribution….
Last week Jess helped me set up the diffusion cell with human albumin and cytochrome c. We ran 3 cells and used membranes from wafer 565. The membranes used were -1,4 (retentate 1), 3,3 (retentate 2), and 4,2 (retentate 3). The gel is posted below: The columns are (from left): Protein, control, retentate 1, filtrate…
Jess showed me how to use the diffusion chamber to separate proteins on the membranes. I ran a gel to see if any proteins appeared in the filtrate. The gel is posted below: The lanes are (from left): protein standard, control, retentate 1, filtrate 1, retentate 2, filtrate 2. It appears that protein diffused through…
I was having trouble using the spectrometer to measure rhodamine concentration in the stir cell. I ran a standardized absorbency of the serial dilution of rhodamine. The curve is posted below. It appears to be linear. The molarity of rhodamine must be above 2uM in order for the spectrometer to read it. I think I…
Lingyun and I used the stir cell and spectrometer to measure the diffusion of rhodamine across the membrane. This was the first time I was able to construct an experiment in the stir cell while keeping both membranes intact. In the past one membrane would rupture or the cell would leak too much. For this…
