Cloning the gene encoding the Human HLA-G protein (25-338aa) into a plasmid vector leads to the formation of recombinant plasmid. The resulting recombination plasmid is transformed into e.coli cells. e.coli cells containing the recombinant plasmid survive in the presence of a specific antibiotic and are selected to be cultured under conditions conducive to the expression of the gene of interest. A N-terminal 6xHis tag is linked to the protein. Following expression, the recombinant Human HLA-G protein is isolated and purified from the cell lysate using affinity purification. Denaturing SDS-PAGE is utilized to resolve the resulting recombinant Human HLA-G protein, demonstrating a purity exceeding 85%.
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