This section provides an overview of the transfer conditions required for performing electrophoretic protein transfer. It also provides some troubleshooting tips for electrophoretic transfer and outlines a general workflow for the protein transfer from gels to membranes. Find procedures for reagent and materials preparation for protein transfer, tank blotting and semi-dry blotting procedures, and microfiltration in the Protocols section below.
Related Topics: Protein Blotting Equipment, Membranes, Transfer Buffers, and Protein Detection and Imaging.
Overall, the procedures and principles for semi-dry and tank transfers are the same. Gels and membranes are pre-wet and equilibrated with transfer buffer, and the gel/membrane sandwich is placed into the transfer apparatus in the correct orientation to ensure transfer of proteins to the membrane. Additionally, the appropriate power conditions must also be selected.
Protein blotting workflow.
Different transfer apparatuses, when used with different gel and buffer systems, require different power settings. The table below provides general guidelines for the voltage and current settings recommended for selected gel and buffer systems. Transfer times are increased for gradient gels and decreased for low molecular weight proteins. The values presented in the table are guidelines — transfer conditions should be optimized for every transfer application. Cooling is generally required for all high-intensity transfers (except when using the Trans-Blot® SD cell) and is recommended for long, unsupervised runs.
Guide to power settings for different gel types.
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