The invention provides a method for preparing a pre-
staining luminescent
protein marker. The method comprises the following steps: acquiring a
protein A
gene, a
protein G
gene, an MBP (
Myelin Basic
Protein)
gene and a
heavy chain constant region gene (CH gene) of a mouse and rabbit derived
antibody, and
cutting and combining the genes according to sizes of different proteins in a pre-
staining luminescent protein marker; expressing the proteins, purifying, and mixing proteins of different molecule weights according to a certain ratio so as to obtain a luminescent protein marker; mixing the prepared luminescent protein marker with the pre-
staining luminescent protein marker according to different ratios. The pre-staining luminescent protein marker prepared by using the method is capable of recognizing primary antibodies or secondary antibodies of different sources, binding sites of proteins are increased, light signals are intensified, not only are general positions of proteins indicated in the
electrophoresis process and after membrane transfer, but also precise positions of the proteins can be indicated on X-
ray films, no extra pre-staining marker channels are needed, no extra antibodies are needed, and the
luminescent proteins can be combined with IgG or anti-rabbit and anti-mouse secondary antibodies derived from species such as human beings, rats, mice and rabbits.