The method of SDS (sodium dodecyl sulfate) polyacrylamide gel electrophoresis for the analysis of Robinia pseudoacacia L. callus was established. To protect proteins in the sample from being proteolyzed, denatured or modified, and to dissolve insoluble proteins, addition of DTT (dithiothreitol) and urea to the final concentration of 0.375mM and 8M respectively was necessary for the sample preparation. Addition of 0.025% of ammonium peroxodisulfate and TEMED (N, N, N', N' -tetramethylethylenediamine) to the separating gel solution and filling the gel at the impregnation speed of 8mm/min. at room temperature was appropriate for the gradient (7.5-15%) separating gel polymerization. Electrophoresis being carried out with an ascending voltage of 80V for the stacking gel and with 120V (2.5 hours) -150V (2.5 hours) -180V(1 hour) for the separating gel efficiently prevented band distortion and protein diffusion.