The invention provides the design, preparation methods and applications of the variant of annexin V. The variant of annexin V is the protein with amino acid residue sequence in sequence 1 in the SEQUENCE LISTING, characterized in that it is the protein derived from sequence 1 by one or several amino acid residues of the sequence 1 being substituted, missing or being added and its amino acid residues have the same activity with the sequence of the sequence 1. The variant protein of the annexin V prepared form the preparation methods of the variant of the annexin V has high purity, high productivity, high labeling efficiency and stability, without effect to the biological function of the variant of the annexin V, which is applicable to industrial production and allows for further research of the variant of the annexin V.