The purpose of this study was to compare four different methods of normalising electromyograms (EMGs) recorded during normal gait. Comparisons were made between the amplitude, intra-individual variability and inter-individual variability of EMGs. Surface EMGs were recorded from the biceps femoris, semitendinosus, vastus lateralis and vastus medialis of ten males and two females while they walked on a treadmill at a self-selected speed. EMGs from the same muscles were subsequently recorded during isometric maximal voluntary contractions (MVCs) and concentric, isokinetic MVCs that were performed between 0.52 and 7.85 rad x s(-1) on a BIODEX dynamometer. EMGs were also recorded during eccentric, isokinetic MVCs between 0.52 and 2.62 rad x s(-1). Gait EMGs were then normalised at 2% intervals of the gait cycle by expressing them as a percentage of the following reference values: the mean (mean dynamic method) and the peak (peak dynamic method) EMG from the intra-individual ensemble average; the EMG from an isometric MVC (isometric MVC method); and the EMG from an isokinetic MVC that occurred with the same muscle action, length and velocity of musculotendinous unit as the gait EMGs (isokinetic MVC method). The isokinetic MVC method produced significantly greater (P<0.05) intra-individual variability compared to the other methods when it was measured using the variance ratio. Inter-individual variability of gait EMGs, again measured using the variance ratio, was also greatest when they were normalised using the isokinetic MVC method. The pattern and amplitude of EMGs normalised using the isometric MVC method and the isokinetic MVC method were very similar (root mean square difference and absolute difference both less than 3%). It was concluded that the isokinetic MVC method should not be adopted by gait researchers or clinicians as it does not reduce intra- or inter-individual variability anymore than existing normalisation methods, nor does it provide a more representative measure of muscle activation during gait than the isometric MVC method.