Preparation of Polyclonal Antibody Against SAX Protein by DNA Immunization

Acta Laser Biology Sinica ›› 2019, Vol. 28 ›› Issue (4) : 368-373.

PDF(1867 KB)
PDF(1867 KB)
Acta Laser Biology Sinica ›› 2019, Vol. 28 ›› Issue (4) : 368-373.

Preparation of Polyclonal Antibody Against SAX Protein by DNA Immunization

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Abstract

Bone morphogenetic protein (BMP) belongs to the cytokine and transforming growth factor beta (TGFβ) superfamily and plays an important role in embryo morphogenesis and organ formation. However, it currently lacks various antibodies to the BMP signaling pathway factor SAX. In this paper, the total RNA was extracted from Drosophila embryos, and the cDNA library was obtained by reverse transcription. The SAX gene fragment was obtained by PCR using the cDNA as a template, and the fragment was ligated into pCAGGSP7 to construct a recombinant plasmid pCAGGSP7sax. BALB/C mice were immunized with plasmid DNA immunoassay to prepare polyclonal antibodies against Drosophila SAX protein. Further analysis showed that the constructed eukaryotic expression recombinant plasmid pCAGGSP7sax DNA had good immunogenicity, and the antiserum antibody titer obtained after immunization of mice reached 1:100. Western blot and Drosophila embryo immunofluorescence assay showed that antiserum can specifically recognize SAX protein. The successful preparation of Drosophila SAX antibody laid the foundation for further study of the role of sax gene and BMP signal in Drosophila heart development.

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Preparation of Polyclonal Antibody Against SAX Protein by DNA Immunization[J]. Acta Laser Biology Sinica. 2019, 28(4): 368-373
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