FAQ
Would a sample that is poorly soluble in rehydration solution cause my gel to have horizontal streaking?
Proteins that are not soluble cannot migrate, and therefore cannot focus. Increase the concentration of the solubilizing components in the rehydration solution.
• Commonly 8 M urea is used, but the concentration can be increased to 9 or 9.8 M if necessary.
• CHAPS, Triton X-100, or NP-40 are the neutral detergents most commonly used. Use at a total combined
concentration not exceeding 4%(v/v).
• DTT or DTE (20 to 100 mM) is the reducing agent most commonly used.
• The recommended IPG Buffer (carrier ampholyte mixture) concentration for the IPGphor system is 0.5%, but up to 2% can be added if sample solubilization remains a problem
What is the composition of IPG Buffer?
IPG Buffers for each pH range are a mixture of carrier ampholytes that enhance sample solubility and produce a more uniform conductivity across the pH gradient during IEF without affecting the shape of the gradient. IPG Buffer can improve separations and sample solubility, particularly with high sample loads. IPG Buffers are also specially formulated to not interfere with silver staining.
How do I choose the pH range of an IPG buffer?
See the Selection guide for IPG buffers