A suitable protein solubilization buffer for 2D analysis must solubilize proteins effectively, without disturbing the native charge of the proteins. Urea based solubilization buffers solubilize proteins effectively, however can modify the native
charge of the proteins, due to carbamylation. Urea exists in equilibrium with ammonium cyanate that modifies α- and ε-amino groups, inducing changes in the isoelectric point of proteins leading to artifactual results.
One way to minimize the risk of carbamylation is to prepare the urea based reagents fresh before each use. G-Biosciences developed 2D-Xtract™, a dry urea based pre-mixed and ready-to-use solubilization buffer.
Simply add an appropriate volume of the supplied rehydration buffer to the dry buffer and then use to solubilize proteins, saving time and improving the quality of IEF/2D gel electrophoresis (Patent Pending).
2D-Xtract™ is also designed to be used as a rehydration buffer for IPG strips.
2D-Xtract™ is based on our successful line of FOCUS™ Extraction Buffers. During the development of the FOCUS™ Extraction Buffers, optimal concentrations of critical agents, buffering and stabilizing agents, including urea, thiourea, CHAPS, and non-detergent sulfobetaines were demonstrated to give optimal protein extraction and improved spot resolution for 2D gel analysis. 2D-Xtract™ has optimized concentrations of urea, thiourea, CHAPS and non detergent sulfobetaine (ND SB) 201.
Convenient and simple to use extraction buffer.
No preparation required, simply hydrate and use.
Prevents urea induced protein carbamylation.
Prevents waste of unused reagents.
No artifactual protein bands due to dust and human skin contamination.