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CUSABIO employed an immunization strategy, immunizing a rabbit with a synthesized peptide derived from human STAT3, to stimulate an immune response. Splenocytes were isolated from the immunized rabbit, and RNA extraction was performed, followed by reverse transcription to convert RNA into complementary DNA (cDNA). Using degenerate primers, the STAT3 antibody gene was amplified from the synthesized cDNA and inserted into an expression vector. The recombinant vector was then transfected into a host system to produce recombinant monoclonal antibodies targeting STAT3. The resulting antibodies were purified using affinity chromatography from the cell culture supernatant. To validate the specificity and affinity of the STAT3 recombinant monoclonal antibody, ELISA, WB, and IHC applications were conducted, demonstrating their ability to specifically recognize human STAT3 protein.
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