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Production and characterization of polyclonal antiserum against DC2 proteinHsu, Chia-wei 09 September 2005 (has links)
The hsDC2, an unknown gene, was located on chromosome 4q25. The genetic protein product contained 149 amino acids with the molecular weight of 16.8 kDa.
By bioinformatics, three predicted non-membrane portion of hsDC2 gene were designed to immune rabbit. The hsDC2 a1~31and hsDC2 a54~82 gene were inserted in pGEX6p-1 that expressed as fusion protein with GST. The other, hsDC2 a140~149, was synthesized with additional cycteine for conjugation with mcKLH at N-terminal. After four times subcutaneous injection, the antiserum was blooded from rabbits. The antibody was purified from the antiserum by protein A beads and confirmed with antigen by western blotting. The identified antibody was used to examine the expression of hsDC2 protein during U937 differentiation induced by RA and PMA.
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