Question

I am needing a product to concentrate 20mg of protein. My current volume is 15mL and I need to get to under 5mL volume. I have used spin columns in the past and my protein has gotten stuck on the cellulose membrane, so I am looking for alternatives.

Poor Quality Detergents Cause Artifactual Results

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Pitfalls of Protein Identification

Proteomics is the analysis of a species' protein complement that is expressed by its genome, but the sheer complexity of the proteome raises significant issues for a researcher.  There are an estimated 20,000-25,000 human protein-coding genes that could code for >1,000,000 proteins, especially when protein modifications are considered. These include N- and C-truncations, alternative spliced forms and the large number of Co- and Post-translational modifications, including:

Question

I am looking to isolate proteins stuck to membranes in human and animal brain. I am then running the protein I retrieve on 2D gels. At the moment I am using a crude method to retrieve these proteins. This method is very time consuming and lacks reproducibility.
Do you have advice for isolating membrane proteins as well as aggregated protein stuck to the membrane for their subsequent analysis by 2D protein electrophoresis? Can I resuspend the proteins straight into my buffer for rehydration and IEF?