Aim: To express the fusion protein of human Nodal mature peptide in prokaryocytes and to prepare mouse anti-Nodal polyclonal antibody.
Methods: The expression vector pReceiver-hNodal was constructed and transformed into BL21(DE3). His(6);-hNodal fusion protein was induced by IPTG and separated on SDS-PAGE and the recovered fusion protein was used to immune BALB/c to produce polyclonal antibody. The titer of the polyclonal antibody was detected by indirect ELISA and its specificity was analyzed byWestern blot and immunochemical staining.
Results: The expected molecular weight of fusion protein around 15 000 was highly expressed in E.coli and specific antibody was obtained.
Conclusion: The successful preparation of anti-Nodal mature peptide polyclonal antibody will lay the basis for further study of the biological function of Nodal on tumor progress.