[Soluble expression, purification of bone morphogenetic protein-2 fragment and its antibody preparation].
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.
Deng
Shao-lin
SL
Chengdu Army General Hospital, Chengdu 610083, China.
Quan
Yi
Y
Pan
Xian-ming
XM
Qu
Bo
B
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism*; D000918:Antibody Specificity; D001483:Base Sequence; D055396:Bone Morphogenetic Protein 2 / Q000276:immunology / Q000302:isolation & purification*; D002460:Cell Line; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000906:Antibodies; D055396:Bone Morphogenetic Protein 2