The analysis of normal vibration mode is a powerful tool, which can not only predict the possible movement of the molecule but also explain more than half of the protein motions by using merely two normal vibration modes of low frequency. The analysis of normal vibration mode is applied broadly in the field of structural molecular biology, such as the structure change of protein, the open and close of molecular ion channel, the movement of ribosome, the surface maturity of virus particle, and so on. The present study uses the “EIN’emo” tool to calculate and analyze the normal mode with low frequency vibration of the molecule, and to get the pattern of frequency and movement calculation. Six isoform proteins are adopted in the study. Furthermore, we get the sequence data of the six sets from the protein data bank, and uploads to the EIN’emo to calculate the low frequency modes of 100 types. The description parameters of frequency and movements are obtained, and the relevance of the frequency and movements is analyzed by the method of the two-dimensional coordinates. Finally, the results of this study provide a practical guide to use the analysis of normal vibration mode effectively to isoform proteins. In the future, we suggest that the analysis of normal vibration mode can be further applied in other research fields.