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Kyunghwa Jeong edited Methods_antibody.tex
almost 9 years ago
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\subsection*{Generation of antibody and western blot}
A polyclonal antibodies against DmCa\textsubscript{v}3 were generated using antigen derived from 302 amino acids at C-terminus of DmCa\textsubscript{v}3
(PCR primers: (primer set for cloning : 5'-GAATTCCAAATTAATCCAATCCGTA-3' and 5'-GCGGCCGCTTAGTCCATGGAGGATT-3').
His-tagged antigen was expressed in \emph{E. coli}, purified and injected into rabbits to generate an immune response (Young In Frontier, South Korea).
We performed our western blot analyses according to standard protocols using rabbit antisera obtained after the third DmCa\textsubscript{v}3 antigen boost. $\beta$-Actin-specific antibodies (Santa Cruz Biotechnology, sc-47778) were used as a loading control.