Diffusion through SC 505
In my last diffusion experiment, I used wafer SC 442. The results seemed to show that IgG was indeed diffusing through the membrane. As a follow up experiment, I am trying to show that I can pass IgG through more than just the one membrane. Furthermore, SC 505 has a much larger average pore size (about 40nm, as compared to 23.5nm in SC 442)
SC 505:
- 5 slots
- average pore size: 39.7nm
- porosity: 5.12%
- cut-off pore size: 49.75 nm
- mean roundness: 0.80, standard deviation: 0.10
Experiment:
1) Buffer A: 55μl of buffer A in well, 10μl of buffer A on top of membranes
2) filtered IgG: 55μl of buffer A in well, 10μl of 1mg/ml filtered IgG on top
Note: The experiment used newly filtered hIgG from a different stock aliquot.
Here are the results from the diffusion experiment (absorption values were found by performing a Bradford assay):
These results are also very promising. The absorption of the filtered IgG filtrate is even higher than the last experiment and the absorption of the filtered IgG stock is lower than that of the stock I used for the first two. See the first experiment here. I would say that we are definitely able to pass IgG through the membranes with diffusion as long as we select wafers with larger pores.
How long was the diffusion run? 1 hour, 24 hours, 48 hours?