[Expression of human HZF1 in E. coli and preparation of antibody against human HZF1 protein].
OBJECTIVE
To express human HZF1 fusion protein in E. coli and to obtain an anti-HZF1 antibody.


METHODS
A DNA fragment encoding non-zinc finger region of HZF1 protein was inserted into pET30a vector to get the recombination expression plasmid pET30a-HZF1. E. coli was transformed with pET30a-HZF1 and the selected clones were cultured with isopropy-beta-D-thiogalactoside induction. The proteins were prepared from the culture and the fusion protein was purified by Ni column. Rabbits were immunized and reinforced three times with the purified fusion protein. The antiserum was collected and the titer and the specificity of the antibody were checked by ELISA and Western blot.


RESULTS
Antibody against HZF1 was obtained and its titer was more than 1:100 000, as proven by ELISA. Western blot analysis showed specific reaction between this antibody and HZF1 fusion protein or the endogenetic HZF1 protein in hemin-induced K562 cells.


CONCLUSIONS
The specific antibody against HZF1 is obtained. The antibody may have potential application in farther HZF1 function study and HZF1 determination in tissues and cells.
Peng
Han
H
National Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005, China.
Zhang
Xin
X
Du
Zhan-Wen
ZW
Zhang
Jun-Wu
JW
D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D018014:Gene Transfer Techniques; D007106:Immune Sera / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014169:Transformation, Bacterial
D004268:DNA-Binding Proteins; D007106:Immune Sera; D011993:Recombinant Fusion Proteins; C514335:ZNF266 protein, human