C.B.S. Scientific EBX-700 - Manual (Page 8)

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8
Instructions for Use
3.1 Blotting Unit Preparation
1. Place 4 freezer blocks in freezer and allow enough time to freeze.
2. Place the blotting chamber on a level work surface in an authorized work area.
3. Consult applications table 4.1 for buffer system and power settings.
4. Place, do NOT drop, a magnetic spin bar (not supplied by C.B.S.) in the bottom of the tank.
This maintains pH, buffer and heat circulation during the electroblot.
5. Place tank on top of the magnetic stirrer.
6. Buffer should be pre-cooled to 10°C.
3.2 Preparing the Gel for Transfer
The EBX-700 can hold up to 4 gel cassettes simultaneously with space provided for a magnetic
stir bar to allow buffer circulation and heat exchange for uniform transfers. When the resolving
gel electrophoresis is complete, proceed with staining and photo-documentation if applicable.
Cut one corner off the gel so that correct orientation is maintained throughout the procedure.
3.3 Preparation of Electroblotting Components
1. CBS membranes are supplied with the blotting paper cut to the correct size for ClearPAGE
gel transfers. If not using CBS membranes, cut transfer membrane to size of gel or sufficient
dimensions to cover the relevant bands. Also, pre-cut Whatman-type 3MM filter paper to
same size and soak in transfer buffer until completely saturated (15 to 30 minutes).
2. Pre-wet membrane chosen according to manufacturer’s instructions. Nylon or Nylon-
supported nitrocellulose should be soaked in ddH
2
O. PVDF in MeOH. Equilibrate all types
in transfer buffer for 3 minutes. Submerge a single foam pad and gently tease to release
trapped air bubbles until pad is completely saturated.
3. Pre-equilibrate gel in 1x transfer buffer to be used prior to electroblotting for 5 to 30 minutes
depending on gel thickness.
4. Open the transfer cassette as shown below and lay it flat on the bench.