[Preparation and characterization of anti-Drosophila Dnop5 antibody].
OBJECTIVE
To prepare the rabbit antibody against Dnop5 and identify its specificity.


METHODS
Dnop5 cDNA was amplified by RT-PCR, and then was subcloned into the fusion expression vectors pET28a(+). After being expressed in E.coli BL21, the truncated Dnop5 protein was purified and used to immunize rabbit. Purified antibody was obtained through affinity chromatography column with the expressed Dnop5. The specificity of the purified antibody was characterized by Western blot and immunohistochemical staining.


RESULTS
The Dnop5 gene was successfully inserted into pET28a(+). After induction, the fusion protein was expressed in the form of inclusion body. The purified fusion protein was obtained by affinity chromatography. After immunization of rabbits, the antibody against Dnop5 was obtained. Western blot analysis and immunohistochemical staining showed that the antibody had a good specificity.


CONCLUSIONS
The rabbit antibody against Dnop5 has been successfully prepared, which lays the foundation for further study on the Dnop5 function.
Yuan
Liu-di
LD
School of Basic Medical Science, Key Laboratory of Developmental Genes and Human Diseases, Ministry of Education, Southeast University, Nanjing 210009, China. yld@seu.edu.cn
Zhang
Yan
Y
Wan
Yong-qi
YQ
Ding
Jie
J
D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000276:immunology*; D000918:Antibody Specificity; D004330:Drosophila; D029721:Drosophila Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D020718:Ribonucleoproteins, Small Nucleolar / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
D000906:Antibodies; D029721:Drosophila Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D020718:Ribonucleoproteins, Small Nucleolar; C499061:nop5 protein, Drosophila