This is the protocol I use to measure band intensities. At the end of the explanation I will post and review my recent results concerning silver stain densitometry. First load a digitized gel into ImageJ Select Image>Type>8 bit to make…

The following changes were made in this adsorption study: The MicroCons/NanoSeps were not spun. We will just characterize the differences between the materials. The pnc-Si membranes were assembled into SepCons because of the potential adsorption to the plastic surfaces. Mass…

Previously I have shown that sample buffer can be used to lift adsorbed proteins off polymer membranes, which can then be imaged on a gel. I have used a similar procedure to assay adsorption on pnc-Si membranes. I used 4…

In anticipation of the pnc-Si separation paper, I have performed a couple of preliminary separations using NanoSeps and MicroCons. I applied 200ul of 1:20 brain extract ( ~.9mg/mL) to 100k NanoSeps and MicroCons, and I spun the samples for 3…

Previously we have studied protein adsorption to pnc-Si membranes by TEM and fluorescence. Neither of these methods will work with polymer membranes. Polymer membranes are too thick to view using TEM. Polymer membranes also autofluoresce creating a lot of noise…

It took me a little while to figure out how to turn the .avi file into a .gif, but here is an image of the diffusion through 501. Due to a problem with MatLab, it took about 5min before I…

I recently set up 24 hour diffusion separations with the BioRad protein standards. The following gel shows the separations using w415 position (-4,3) and w416 (-1,3). Different cutoffs were observed with the two membranes.

We’ve recently put together our own protein ladder, which contains myosin heavy chain, beta-galactosidase, phosphorylase b, IgG, albumin, and cytochrome c. This is my first attempt running this ladder on w398, one of the new pinhole free membranes. From the…

UPDATE:Chris took the membrane sample and normalized with by the no spanning membrane sample. This was plotted against the theoretical spectrum of single crystal silicon. The curves have a similar shape, but the single crystal silicon has lower transmission. This…