[Preparation and characterization of rabbit antibody against human selenoprotein P].
OBJECTIVE
To prepare rabbit antibody against human selenoprotein P (HSelP) by using HSelP expressed in the prokaryotic expression system and identify its specificity.


METHODS
HSelP gene fragment was expressed in E.Coli via IPTG induction and purified through DEAE and Ni-NTA columns sequentially. The purified HSelP as immunogen was used to immunize rabbit. The specificity of rabbit anti-HSelP antibody was analyzed by Western blot.


RESULTS
HSelP protein had been expressed and purified successfully. The polyclonal anti-HSelP antibody could specifically recognize natural HSelP from human serum.


CONCLUSIONS
Polyclonal anti-HSelP antibody with high specificity and purity has been prepared by using purified HSelP as immunogen, which lays the foundation for further research on the function and distribution of HSelP, as well as developing a simple method for detecting HSelP.
Fang
Qing
Q
National Laboratory of Molecular Oncology, Cancer Institute, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100021, China. qingfang71@yahoo.com
Zhang
Ming-cheng
MC
Zha
Yuan-yuan
YY
Bi
Sheng-li
SL
Li
Zhi-wei
ZW
Yang
Jian-guo
JG
Xiao
Zhu-xiu
ZX
Lin
Chen
C
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D018076:DNA, Complementary / Q000032:analysis; D006801:Humans; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011817:Rabbits; D051149:Selenoprotein P; D051140:Selenoproteins
D000906:Antibodies; D018076:DNA, Complementary; D011506:Proteins; D051149:Selenoprotein P; D051140:Selenoproteins