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Sample Preparation Solutions for Cellular Protein Fractionation Poster, Rev A

E. coli samples were treated using a carbonate extraction procedure to extract and enrich for membrane proteins (Figure 2). Table 2 shows that many of the proteins that were extracted using this procedure are membrane-associated E. coli proteins.


Conclusions
Applying specialized sample preparation solutions for the fractionation of cellular proteins from complex protein mixtures provides a simple and rapid method for enrichment of both lowabundance and membrane-associated proteins prior to sample analysis by 2-D electrophoresis. Understanding the cell biology of membrane proteins is especially important in the search for new drug targets, as ~50% of the known drug targets are membrane proteins. We have shown that application of four new kits for protein fractionation can provide a simple way to differentiate the proteome of a complex sample into a number of manageable fractions. Key features of these kits include:

Rapid separations Extraction from a crude sample to an enriched, gel-ready sample takes <3 hr

Reproducible results Each kit is based on proven chemistries and provides detailed instructions for application

Increased protein solubilization The PSB solution has been optimized to enhance solubilization of membrane-associated proteins for 2-D separation

Improved 2-D results The ReadyPrep 2-D cleanup kit provides quantitative recovery of samples with elimination of detergent and salt contaminants

Applying these sample prep kits to enrich proteins from complex biological samples should enhance the quality of the data that is obtained with any 2-D proteomics program.


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