The Protein Man's Blog | A Discussion of Protein Research

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?

Protein Research Requires Protein Estimation;
But What is the Best Protein Assay?