Native gels rely on alkaline pH (instead of SDS) to give proteins a negative charge, causing it to migrate towards the positive anode.

Keeping proteins in native configurations means not heating the sample, and using a glycerol-based sample buffer instead of a urea based sample buffer.

ß-mercaptoethanol can be added to sample buffer to reduce disulfide bonds

Gel buffer 1.5M Tris-HCl pH 8.8

2x sample buffer 2x Tris-glycine pH 8.3 (0.192MGlycine, 0.025M Tris)
0.02% bromophenol blue
20% Glycerol for native conditions
(or 4M urea for denaturing conditions)

1x running buffer 0.192M Glycine, 0.025M Tris pH 8.3

Pour 1.5mm separating acrylamide gel as in SDS-PAGE, but omit SDS from gel. Gel buffer is 4x for separating gel.

Pour stacking gel as in SDS-PAGE, but omit SDS. Gel buffer is 12x for stacking gel.

Run gel in cold room to prevent diffusion. Use low voltage (40-50V) to prevent smearing of bands.