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The ATG12 recombinant monoclonal antibody is produced using DNA recombinant technology and in vitro genetic manipulation. The process involves immunizing an animal with a synthesized peptide derived from human ATG12, isolating B cells, and selecting positive B cells. These selected B cells undergo screening and single clone identification. The genes encoding the light and heavy chains of the ATG12 antibody are amplified via PCR and inserted into a plasmid vector to create a recombinant vector. This vector is then transfected into a host cell line for antibody expression. The ATG12 recombinant monoclonal antibody is purified from the cell culture supernatant using affinity chromatography. It exhibits specificity for human ATG12 protein and finds application in ELISA.
The ATG12 protein is a key component of the autophagy pathway in cells. In the autophagy pathway, ATG12 forms a conjugate with ATG5 and ATG16L to form the ATG12-ATG5-ATG16L complex, which acts as an E3 ligase, catalyzing the lipidation of LC3. The lipidated form of LC3, called LC3-II, is recruited to the autophagosomal membrane, where it helps to form the autophagosome. The autophagosome then fuses with lysosomes, where the contents are degraded and recycled.
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