I'm trying to implement an electrophoretic technique to detect peptides from casein digestion. I was thinking of Tris-Tricine since is the usually employed for peptides of lower molecular weights, but the techniques always mention the use of both buffers. In the lab, we use a Mini Protean tetra system electrophoresis chamber.
Dear Manuel, I used Tris-Tricine electrophoresis following the original protocol invented by Schagger and von Jagov in 1987 (see Anal. Biochem,,166, pp.368-377). The use of high concentration separating gels (16.5%) and TWO ELECTRODE BUFFERS (cathode and anode) is critical to get small proteins/peptides separated. In some cases, adding urea to the separating gel buffer also helps. In any case, this method gives more or less good resolution for peptides higher than 1.5 kD, and, if you tend to stick with tricine-method, I would recommend to try first some standard peptides. Good luck
In addition. I used Miniprotean II system, there should not be any problem, but definitely, if you used a larger gel you would get a better resolution. Also, you need to find a peptide map for casein (using which protease?-trypsin) to see how many large peptides if any you may expect. Best.
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