Buffer solution reagent FOCUS™

for electrophoresisproteinurea

One of the most important considerations before running 2D gel electrophoresis is the choice of protein solubilization buffers. A suitable buffer must solubilize proteins effectively, without disturbing the native charge of the proteins. Urea, a common chaotrope, is widely used for solubilization and denaturation of proteins. One of the disadvantages of using urea is carbamylation. In water, urea exists in equilibrium with ammonium cyanate, the level of which increases with increasing temperature and pH. Cyanate reacts with α-amino and ε-amino groups of proteins and induces a change in the isoelectric point of proteins. This leads to artifactual results. Therefore, carbamylation must be avoided. One way to minimize the risk of carbamylation is to prepare the urea-based reagents fresh before each use. G-Biosciences has developed a series of dry, urea-based, pre-mixed and ready-to-use solubilization buffers. Simply add an appropriate volume of the supplied rehydration buffer to the dry buffer. Then use to solubilize proteins, saving time and improving the quality of IEF/2D gel electrophoresis (Patent Pending). FOCUS™ Extraction Buffers are also designed to be used as rehydration buffers for IPG strips. FOCUS™ Extraction Buffers are experimentally optimized concentrations of critical, buffering and stabilizing agents, including urea, thiourea, Nonidet® P-40, CHAPS and sulfobetaines. The FOCUS™ Extraction Buffers are designed to produce optimal protein extraction and improved spot resolution for 2D gel analysis.