Rapid Transfer Buffer (10X) (Powder)

Description: Quick, efficient transfer of proteins from SDS-PAGE gels to membranes for Western blotting applications Directions for Use : Stock Solution: 1L size: Dissolve the powder in 1L deionized water for Rapid Transfer Buffer (10X). 100 ml size: Dissolve the powder in 100 ml deionized water for Rapid Transfer Buffer (10X). Working Solution: Prepare 1 L of Rapid Transfer Buffer (1X) by diluting 100 mL of Rapid Transfer Buffer (10X) with 800 mL of deionized water, then add 100 ml Ethanol.Application : Wet Transfer Protocol 1. Prepare membrane and filter paper for transfer: a. A blotting membrane and 3 pieces of filter paper should be cut to fit dimensions of the gel. Note: PVDF membranes must be pre-wetted according to the manufacturer’s instructions in 100% methanol prior to equilibration in transfer buffer. b. Equilibrate the membrane and filter paper in 1X Rapid Transfer Buffer for a minimum of 5 minutes. 2. Following protein electrophoresis, assemble the blotting sandwich following the manufacturer’s instructions for the transfer apparatus. 3. Place the blotting sandwich in a wet transfer tank filled with 1X Rapid Transfer Buffer. 4. Transfer constant current 300 - 400 mA at room temperature. Semi-Dry Transfer Protocol 1. Prepare membrane and filter paper for transfer: a. A blotting membrane and 3 pieces of filter paper should be cut to fit dimensions of the gel. Note: PVDF membranes must be pre-wetted according to the manufacturer’s instructions in 100% methanol prior to equilibration in transfer buffer. b. Equilibrate the membrane and filter paper in 1X Rapid Transfer Buffer for a minimum of 5 minutes. 2. Following electrophoresis, wash the gel for 2 minutes in deionized water. 3. Pre-equilibrate the gel in 1X Rapid Transfer Buffer for 5 minutes. 4. Assemble the blotting sandwich following the manufacturer’s instructions for the semi-dry transfer apparatus. 5. Transfer constant current 300 - 400 mA at room temperature.Storage/Stability : Store at 25°C for 2 years.