Preparation and identification of the Syntenin1 polyclonal antibody, a cancer metastasis related gene

To express the GST fusion protein, GST-Syntenin1 in E. coli, and to prepare the polyclonal antibody of Syntenin1. CDS fragment of Syntenin1 was obtained by RT-PCR from normal mouse brain and subcloned into pGEX-4T-2 to generate pGEX-4T-2-Syntenin1 recombinant. The confirmed recombinant was transform...

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Published inSichuan da xue xue bao. Journal of Sichuan University. Yi xue ban Vol. 40; no. 6; p. 1119
Main Authors Xia, Xiao-Qiang, Chen, Mi-Na, Li, Fang-Hua, Cai, Pei-Ling
Format Journal Article
LanguageChinese
Published China 01.11.2009
Subjects
Animals
Antibodies - immunology
Antibodies - metabolism
Escherichia coli - genetics
Escherichia coli - metabolism
Genes, Neoplasm - immunology
Glutathione Transferase - biosynthesis
Glutathione Transferase - genetics
Humans
Rabbits
Recombinant Fusion Proteins - biosynthesis
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - immunology
Syntenins - biosynthesis
Syntenins - genetics
Syntenins - immunology
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Summary:To express the GST fusion protein, GST-Syntenin1 in E. coli, and to prepare the polyclonal antibody of Syntenin1. CDS fragment of Syntenin1 was obtained by RT-PCR from normal mouse brain and subcloned into pGEX-4T-2 to generate pGEX-4T-2-Syntenin1 recombinant. The confirmed recombinant was transformed into the BL21 competent cells and induced with IPTG. The recombinant fusion protein was purified with immobilized Glutathione Sepharose and confirmed by SDS-PAGE. The purified fusion protein was mixed with the Freund's adjuvant, and then injected into New Zealand white rabbits by hypodermic injection. The polyclonal antibody titer and specification were identified by Western blot. Syntenin1 polyclonal antibody bind Sytenin1 protein specifically and the antiserum tiger reached to 1 : 20 000. The Syntenin1 polyclonal antibody with high titer and high specificity was prepared successfully. This will be very helpful for the further study on Syntenin1 function and molecule mechanism of cancer metastasis.
ISSN:1672-173X