[Preparation and identification of the Syntenin1 polyclonal antibody, a cancer metastasis related gene].
OBJECTIVE
To express the GST fusion protein, GST-Syntenin1 in E. coli, and to prepare the polyclonal antibody of Syntenin1.


METHODS
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.


RESULTS
Syntenin1 polyclonal antibody bind Sytenin1 protein specifically and the antiserum tiger reached to 1 : 20 000.


CONCLUSIONS
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.
Xia
Xiao-Qiang
XQ
Laboratory of Molecular Neurobiology, State Key Laboratory of the Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
Chen
Mi-Na
MN
Li
Fang-Hua
FH
Cai
Pei-Ling
PL
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D052138:Genes, Neoplasm / Q000276:immunology; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D053676:Syntenins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
D000906:Antibodies; D011993:Recombinant Fusion Proteins; C109621:SDCBP protein, human; D053676:Syntenins; D005982:Glutathione Transferase