This past week I ran multiple experiments using the pressure cell and sep-cons. I used membranes from W684 for the sep-cons since the burst pressure of these memebranes is high (around 3.65-10Psi). I added 100uL BSA to the Sep-Con and put 20uL 1xPBS on the backside. Before adding the NC to the sep-con, I assembled…
I’m having trouble getting the PDMS to cure on the NC. When I first started these experiments in August, the PDMS seemed to cure on the NC without a problem. Now, when I remove the PDMS + NC from the oven, after heating for 2+ hours at the highest setting, I notice that the PDMS…
Creating an array to hold a pnc-Si wafer to proving to be difficult. The PDMS spreads too rapidly though the NC to form a consistent array for the pnc-Si membrane (and eventually wafer) to rest on. I’m still running different trials of the PDMS on NC to see if I can get the array to…
What has been accomplished thus far: Able to detect BSA on NC membrane with the use of Ponceau S stain With PDMS array on NC, no BSA leaks through the area covered by PDMS Initial trial done with 10:1 ratio of elastomer base: curing agent. Later 3:1 and 5:1 ratios done. 3:1 is too sticky…
During the past week I tried various methods of applying PDMS to nitrocellulose. Below is a table of what the PDMS array looks like on the NC before curing in the over, and after applying 1mg/mL BSA and staining with Ponceau S. When I painted the PDMS on, I used the thin side of the…
I ran a few experiments using the stir cell on the shaker table. Diffusion occurs faster on the shaker, but more air bubbles form around the membrane, thus preventing further passage of rhodamine. The first experiment I ran was with .2mM rhodamine diluted from 2mM in a PBS solution. For all of the experiments PBS…
Below is an image of the PDMS array on the nitrocellulose membrane. I’m trying to create a stamp that can be applied to the NC membrane, before PDMS is poured on. The idea is that the stamp will be heavy enough to keep the PDMS from soaking into the areas of the NC that the…
I’m trying to find the best method to stain proteins on nitrocellulose, so we can easily see the protein on the NC membrane. I first tried Coomassie Blue which did not work very well with the NC. The Coomassie was too dark, as seen by the image below. Harold Smith gave me some Ponceau S…
Last week I tried to diffuse Cytochrome C through our membranes onto PVDF and PES membranes to see if we can detect protein adsorption on the other membranes. The trial illustrated that cytochrome c will diffuse through our membranes and stick to the PVDF and PES membranes. I ran the experiment again, but I used…
Harold Smith at the Med Center proposed an experiment for Jess and I to try, in which we use our membranes to separate proteins onto a PES, PVDF (Polyvinylidene fluoride) or Nitrocellulose membrane, similar to a Western Blot of DNA microarray. The idea is to create a system to screen for drugs that break protein-protein…