Sauer:bis-Tris SDS-PAGE, the very best: Difference between revisions

You can use any of the the common Laemmli-like buffers. Many people over-cook their proteins and cause brakdown, probably because Laemmli was solubilizing a protein pellet. A short heating of ~1 min at ~80-100 C is more than sufficient and only has to be done once to fully denature the proteins and coat them. Using a lower temp, like 40-60 C to thaw frozen aliquots helps to prevent ruining the samples.  

Store this concentrate and add 2-mercaptoethanol to ~1% (~140 mM) to an aliquot within a few days of use. There is a lot of 2-ME in many recipes, prbably because of its short half-life (compensating for decay). There is no way to know the reducing activity in older preprarations; adding fresh to 1% a few times during the use of the aliquot is a cheat, but works fine for most applications.