Method for preparing pre-staining luminescent protein marker

Abstract

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.

Description

technical field [0001] The invention relates to a preparation method of a pre-stained luminescent protein marker, which belongs to the fields of molecular biology and genetic engineering. Background technique [0002] Protein markers are widely used in protein electrophoresis as a standard for protein molecular weight. It is a mixture of known proteins or polypeptides with different molecular weights. Western Blot needs a ruler for tracking the detection of positive antigen signals and transmembrane efficiency, so the pre-stained protein Marker was produced. Pre-stained protein markers are formed by chemically modifying each protein (or polypeptide) in a common protein marker and covalently coupling it with a blue or other colored dye. The appearance of pre-stained protein markers has brought great convenience to Western Blot. After transferring to the membrane, the researchers can determine the approximate position of the target band according to the size of the blue (or ...

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Problems solved by technology

[0005] However, the current products have certain deficiencies. The biotinylated protein marker needs to be added with anti-biotin-HRP antibody or streptavidin-HRP, which makes the experimental operation cumbersome.

In addition, only luminescent protein markers cannot track and detect positive antigen signals and membrane transfer efficiency. An additional pre-stained marker is required, which is not only cumbersome but also takes up swimming lanes, reducing the number of samples that can be electrophoresed

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