Preparation method of luminescent protein marker

Abstract

The invention provides a preparation method of a pre-dyed luminescent protein marker. The method comprises the steps of obtaining a protein A gene, a protein G gene, an MBP gene, and a heavy chain constant region gene (CH gene) of mouse and rabbit source antibodies, and cutting or combining the genes according to the sizes of various proteins in the pre-dyed luminescent protein marker; and expressing the proteins, purifying the proteins, and mixing the proteins of different molecular weights at a certain ratio to obtain the luminescent protein marker. The luminescent protein marker is capable of identifying a first antibody or a second antibody of different sources, increasing the binding sites of the proteins and thus strengthening an optical signal. The luminescent protein marker can be exposed and displayed on an X film after being incubated together with the first antibody or the second antibody without an extra antibody; the luminescent protein can be combined with IgG from species, such as human, rat, mouse and rabbit or anti-rabbit, and anti-mouse second antibodies; and compared with luminescent markers of other companies, the pre-dyed luminescent protein marker has a wider indication range and has the molecular weight ranging from 16kDa to 200kDa.

Description

technical field [0001] The invention relates to a method for preparing a photoprotein 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. Protein markers are divided into ordinary markers, pre-stained markers and luminescent markers. Ordinary marker bands can only be seen after Coomassie brilliant blue staining, so they can only be used for [0003] In SDS-PAGE electrophoresis. 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 ...

AI Technical Summary

Problems solved by technology

[0006] 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.

ProteinA and ProteinG are selective for the binding of different species of antibodies, resulting in some antibodies that cannot be displayed on X-film

Moreover, most of the currently available markers have a limited indication range, and cannot clearly indicate large molecular weight proteins.

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