Semi-dry protein transfer
Semi-Dry Protein Transfer
- Cut the transfer membrane and four sheets of Whatmann 3 MM paper on gel size (7 x 8 cm).
- Equilibrate the membrane in transfer buffer (Towbin buffer, cat. no. 42558) or, if PVDF membranes are used, first in ethanol for 2 minutes and then in transfer buffer for another 5 minutes.
- Also moisten the four sheets of Whatmann 3 MM paper with transfer buffer.
- Remove the gel from the cassette and equilibrate the gel in transfer buffer for 5 minutes.
- Build the transfer sandwich analogously according to the manufacturer's instructions for the blotter and place it in the semi-dry blotter.
- The transfer takes place at room temperature with 1.5 mA / cm2 gel area for about 1 hour (for standard marker proteins).
When transferring proteins of different sizes, it is advisable to use a discontinuous blotting buffer system (SERVA Semi-Dry Blotting Kit, cat no. 42559.01).