Extraction Buffer for 2D Gel with Diluent, 50ml Urea and NP-40: 2D buffers must solubilize proteins effectively without disturbing the native charge on the proteins. Urea, a common chaotrope, is widely used for solubilization and denaturation of proteins.
One of the disadvantages of using urea is carbamylation. Urea in water exists in equilibrium with ammonium cyanate, of whose level 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. This leads to artifacts in results and therefore carbamylation must be avoided.
One way to minimize the risk of carbamylation is to prepare the urea based reagents fresh before each use. Bio Basic Inc. has developed a series of dry urea based pre-mixed and ready-to-use solubilization buffers. Simply add an appropriate volume of the supplied diluent buffer to the dry powder and then use directly to solubilize proteins, saving time and improving the quality of IEF/2D gel electrophoresis.
2D Sample Extraction Buffers are also designed to be used as rehydration buffers for IPG strips. 2D Sample Extraction Buffers are experimentally optimized with proprietary technologies and critical buffering and stabilizing agents, including urea, thiourea, Nonidet® P-40, CHAPS, and sulfobetaines.
Our 2D Sample Extraction Buffers are designed to produce optimal protein extraction and significantly improved spot resolution for 2D gel analysis. A range of 2D Sample Extraction Buffers have been developed and depending on the nature of the samples, one or more of the buffers suitable for your applications can be ordered.
PL037 is suitable for most applications, however for stronger solubilization effects, we recommend PL038-PL042.