This is a summary of diffusion experiments with A3G that were conducted during the week of 5/10/2010-5/14/2010. The HMM structure of A3G is a large protein multimer held together with RNA. Since the ultimate goal of this project is to…

These experiments were conducted during the week of 5/10/2010 – 5/14/2010. To recap on this project, Barrett and I have been working with Dr. Harold Smith in the hopes of developing a drug screening test that consists of small-scale diffusions.…

I recently began working directly with Dr. Harold Smith’s A3G protein. In my previous posts,  I worked with BSA and IgG in order to develop a small-scale diffusion set-up able to reach equilibrium before it dries out. These preliminary steps…

During the past few weeks, I have been experimenting with different array setups for diffusion. I have been working with BSA, but I will move to APOBEC3G protein once the arrays Barrett designed are finished. All of the pnc-Si arrays…

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…

The results of my last diffusion experiment regarding BSA and filtered IgG (using SC 364) were inconclusive. (See “Diffusion Test– filtered IgG and BSA”) Looking at the histogram of results, there is no clear difference between the absorption of filtered…

The human IgG did not seem to diffuse through the membrane the first time we conducted a diffusion experiment. Barrett believed that the lack of diffusion was most likely due to protein aggregates blocking the pores. So, the human IgG…

SepCon Experiment (Carrie): Does it work in a SepCon format? Use “Protein A Beads and IgG Binding and Filtering (with SepCon Format)” 9/29/09 Make Buffer B at pH 3 9/30/09 1) Rinse off ethanol 2) Add IgG Take absorbance reading…

Test of Binding/ Unbinding (Carrie and Barrett): Test to see if we can successfully bind Human IgG to the protein A beads and then remove it with buffer B. After ordering the new human IgG and protein A beads, I…

Last week, Barrett and I made up the three buffers needed to perform this test: Dissociating buffer: 0.1 M glycine-HCl, pH 3.5 1 M Tris Binding buffer: 0.1 M sodium phosphate, 0.15 M NaCl, pH 7.5 In addition, I made…