Tank blotting

Tank Blotting

Transfer buffer
(10x concentrate):

0.25 M Tris
1.92 M Glycin

Working solution: Dilute 100 ml of 10x concentrate with  200 ml methanol and
700 ml distilled water.


  1. Cut transfer membrane and four pieces Whatmann 3 MM paper to gel size (7 x 8 cm).

  2. Equilibrate the membrane in transfer buffer (Towbin buffer, cat. no. 42558). By use of PVDF membranes equilibrate the membrane first for 2 minutes in methanol and then for additional 5 minutes in transfer buffer.

  3. Wet the porous pads as well as the four pieces Whatmann 3 MM paper with transfer buffer.

  4. Remove the gel from the cassette and equilibrate the gel for 5 minutes in transfer buffer.
  5. Mount the transfer sandwich according to the manufacture’s protocol and place it in the tank blotter.

  6. Transfer is done at room temperature at 250 mA resp. ca. 60 V for ca. 1 - 2 hours.

After blotting proteins can be stained on the membrane:

  • Detection with Ponceau S solution (0.2 %, cat. no. 33427)
  • Staining with Amido black

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