Hello. That does not seem an easy task, if you would like to use the classic Tris/Glycine system. The problem is that the process in the stacking gel (called isotachophoresis and responsible for the stacking) takes place at pH of 6.8. One possibility would be to search for an appropriate buffer system, different from Tris/Gly (unfortunately I have never ask myself for such one for the respective pH, one possible suggestion could be found here: https://www.thermofisher.com/de/de/home/life-science/protein-biology/protein-gel-electrophoresis/protein-gels/nupage-protein-gels/nupage-tris-acetate-gels.html). Another option would be to try continous electrophoresis without stacking gel, which will not give you a sharp bands, but your protein will remain intact.

Hello. That does not seem an easy task, if you would like to use the classic Tris/Glycine system. The problem is that the process in the stacking gel (called isotachophoresis and responsible for the stacking) takes place at pH of 6.8. One possibility would be to search for an appropriate buffer system, different from Tris/Gly (unfortunately I have never ask myself for such one for the respective pH, possible suggestion could be a Tris/acetate gel). Another option would be to try continous electrophoresis without stacking gel, which will not give you a sharp bands, but your protein will remain intact.

Hi there,

If you are able to load very small volumes of samples you may try to run SDS PAGE without any stacking gel.

What about gradient gels? The gradient achieves an effect similar to stacking, but without the pH transition. They're commercially available, and they can be hand cast, though the technique for doing so is beyond my experience.

Just a clarification sought-do you need to run the SDS-PAGE i.e the denaturing PAGE for resolving your protein of interest from a mixture or is it the native PAGE i.e tto resolve the intact protein in its native form (without disturbance to the PTM you have mentioned)? If it is the latter, dilute the 1.5 M Tris .Cl (pH 8.8) to get a final concentration of 0.5 M TrisCl pH 8.8 (instead of 6.8) & use the same volume that you would use for your usual stacking gel & run your samples (devoid of SDS, b-mercaptoethanol etc).