Bio-Rad Trans-Blot® Turbo™ Transfer System User Manual

Page 27

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3.7 Transfer Using Traditional Semi-Dry Consumables

Typical procedure utilizing the Trans-Blot Turbo system and conventional semi-dry western blotting
consumables is detailed below:

1. Equilibrate the gel in Towbin transfer buffer (25 mM Tris, 192 mM glycine pH 8.3, 20% MeOH) for 10


2. Soak two pieces of extra-thick (2.4 mm) filter paper in transfer buffer. Six pieces of thick (0.8 mm)

filter paper can be used if extra-thick paper is not available.

3. While the gel is equilibrating, prepare a transfer membrane. Wet a nitrocellulose membrane briefly

in transfer buffer or PVDF membrane in methanol or ethanol for 30 sec, then wash in water for 1–2
min, and equilibrate in transfer buffer for at least 10 min with agitation.

4. Assemble the transfer sandwich on the cassette base (anode) by placing one piece of wet extra-

thick or 3 pieces of thick filter paper on the bottom, then the membrane, the gel, and finally,
the remainder of the wet filter paper on top. Use the blot roller to remove air from between the
assembled layers (Figure 19).

5. Once the stacks are positioned in the cassette base, place the cassette lid on the base. The lid is

reversible, but ensure that the electrical contacts fit closely into the slots in the base. Press the lid
down firmly and turn the dial clockwise to engage the lid pins into the locking slots.

6. Load a second cassette if desired. Refer to Table 1 for the appropriate combinations of gels that

can be combined in a single run.

7. Slide the cassette (with the dial facing up) into the bay until it makes contact with the magnetic

interlock and you hear a click. Cassettes can be inserted into the bays in any order, with or without
power to the system.

8. Select the LIST button from the Home menu and the STANDARD SD transfer protocol from the

Bio-Rad preprogrammed protocols or the user-defined protocol of choice.

9. To initiate the run, press the navigation button that corresponds to A:RUN for the cassette in the

upper bay or B:RUN for the cassette in the lower bay.

For futher information, refer to the Bio-Rad Protein Blotting Guide: A Guide to Protein Detection and
Blotting, bulletin 2895, for detailed information. Bulletin 2895 can be downloaded from our website,

, as a PDF file, or contact Technical Support at 1-800-424-6723.

Fig. 19. Proper assembly of blotting sandwich using traditional


Top (

) cassette

electrode (cathode)

Top ion reservoir


Blotting membrane

Bottom ion reservoir




) cassette

electrode (anode)


Tech Support: 1-800-424-6723 • 15

Trans-Blot® Turbo™ Blotting System