Protein Protocols Protein Protocols

340 Delcuve and Davie
3. The transfer efficiency of basic proteins from AUT minislab polyacrylamide gels to nitrocellulose
membranes was poor when a Tris-glycine-methanol (25 mM Tris, 192 mM glycine,
20% [v/v] methanol, and 0.1% SDS) transfer buffer was used.
4. With the Bio-Rad Trans-Blot cassette, four minislab gels can be easily accommodated.
5. Following transfer and baking, the nitrocellulose membrane may be stored for several
weeks at room temperature before proceeding with immunochemical staining.
6. Leaving the nitrocellulose membrane in water for too long after staining with India ink
will result in removal of the stain.
7. We have used this alkaline transfer buffer to transfer histones from SDS slab gels to nitrocellulose
membranes. Pretreatment of the SDS slab gel is not required. However, we have
found that washing the SDS slab gel in equilibration buffer 2 for 30 min improved the
efficiency of elution of the histones from the SDS gel.
Acknowledgments
This work was supported by grants from the Medical Research Council of Canada
(MT-9186, MT-12147, MA-12283, PG-12809) and the University of Manitoba Research
Development Fund.
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