Optimizing Transfer Conditions - BIO RAD Trans-Blot Turbo Instruction Manual

3.8 Optimizing Transfer Conditions

The following techniques, alone or in combination, will increase transfer efficiency:
• Use a low-percentage gel or a gradient gel. High-percentage gels retard protein transfer, especially
with large proteins
• High molecular weight proteins may require increased transfer times, particularly when using thick
gels. Increase the transfer time or power conditions
• With long transfer times or high power conditions, some very low molecular weight proteins may
transfer through the membrane to the lower ion reservoir stack. Use a shorter transfer time or
reduce power conditions for most effecient transfers
• Use the blot roller to remove any air bubbles when assembling the transfer sandwich. Air bubbles
between layers of the assembled sandwich will prevent protein transfer, producing blank spots on
the membrane.
Table 4. Recommended power conditions for transfer using the Trans-Blot Turbo system.
With Turbo Transfer Packs
With Standard Semi Dry Components
Refer to the Bio-Rad Protein Blotting Guide, bulletin 2895, for more information on optimizing
electrophoretic transfer. The Protein Blotting Guide can be downloaded from our website,
www.bio-rad.com, as a PDF file, or call Technical Support at 1-800-424-6723.
Trans-Blot ® Turbo™ Blotting System
16
Single Mini Gel
25 V, 1.3 A, 7 min
25 V, 1.0 A, 30 min
Two Mini Gels or One Midi Gel
25 V, 2.5 A, 7 min