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[Soluble expression, purification of bone morphogenetic protein-2 fragment and its antibody preparation].

Authors :
Deng SL
Quan Y
Pan XM
Qu B
Source :
Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition [Sichuan Da Xue Xue Bao Yi Xue Ban] 2008 Jan; Vol. 39 (1), pp. 19-22, 51.
Publication Year :
2008

Abstract

The bone morphogenetic protein-2 (BMP-2) fragment (BMP-2omega, 606-846bp) cDNA was amplified from total RNA of SAOS-2 cells by using RT-PCR. The PCR product was then inserted into pET-28a (+) vector for constructing the expression plasmid that would be used to transform the host cell BL21(DE3). After IPTG inducing under different conditions, this BMP-2w protein could be expressed in high level as a soluble form, and purified by chelating column (Ni-NTA). Polyclonal antibody was made by immunizing mice with using purified protein, and the antiserum titer generated was 1: 6400 that was measured by ELISA. Western blot result showed that this antibody could bind to BMP-2 protein specifically. Above research result set up the basis for studying on the treatment of osteosarcoma.

Details

Language :
Chinese
ISSN :
1672-173X
Volume :
39
Issue :
1
Database :
MEDLINE
Journal :
Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition
Publication Type :
Academic Journal
Accession number :
18390191