To make 1L of 1X transfer buffer: Mix 100 ml of 10X transfer buffer, 200 ml of methanol, and 700 ml of ddH2O and store at 4ºC for up to one week. This product supplies enough 10X material to make 10 liters of 1X solution. Generally, 20% methanol is recommended, however it may be beneficial to decrease methanol concentration to 5-10% for increased transfer efficiency of large, low abundancy proteins. CST recommends electrotransferring to 0.2 μm pore size nitrocellulose membranes at 70 volts for 2 hours.
Store 10X buffer at room temperature. At 10X, this buffer is stable for 24 months. Store at 4°C and use within 1 week once it has been diluted to 1X and methanol is added.
Tris-Glycine Transfer Buffer (10X) is used as a transfer buffer during western blotting. Product is shipped and stored at room temperature.
1X Formulation: 25 mM Tris, 192 mM Glycine, 20% (v/v) methanol, pH ~8.3.
Note: Methanol is not supplied but is required.
Tris-Glycine Transfer Buffer (10X) is a commonly used western blot buffer for the electrotransfer of proteins from SDS-PAGE gels to nitrocellulose or PVDF membranes. The formulation is based on the widely accepted Towbin transfer buffer (1) and is for use in tank (wet) transfer systems, the recommended system used by Cell Signaling Technology (CST) scientists.