Preparation of antibodies against a novel leucine-zipper protein, Ecp.
- Author:
Dao-Yong WANG
1
;
Li LIU
;
Wei XIE
Author Information
1. Genetics Research Center, Department of Genetics and Developmental Biology, Southeast University, Nanjing 210009, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies;
immunology;
isolation & purification;
metabolism;
Blotting, Western;
Chromatography, Affinity;
Drosophila Proteins;
genetics;
immunology;
Drosophila melanogaster;
growth & development;
metabolism;
Electrophoresis, Polyacrylamide Gel;
Gene Expression Regulation, Developmental;
genetics;
physiology;
Green Fluorescent Proteins;
genetics;
metabolism;
Peptide Initiation Factors;
genetics;
immunology;
Polymerase Chain Reaction;
Rabbits;
Recombinant Fusion Proteins;
genetics;
immunology;
metabolism
- From:
Chinese Journal of Biotechnology
2003;19(3):291-293
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this research is to prepare high quality polyclonal antibodies against Ecp, a recently identified leucine-zipper protein. The full length cDNA of ecp was amplified by PCR, cloned into pGEX-4T-1(His)6 and transformed into E. coli DH 5alpha. After induction with IPTG, the GST-Ecp fusion protein from the lysate was bound to glutathione-Sepharose 4B and digested with thrombin. The released Ecp protein was further purified through Ni-NTA affinity chromatography to homogeneity. A rabbit was immunized with the purified Ecp, and the antibody generated against Ecp was purified by affinity chromatography. The results of the Western blot showed that Ecp is present in various development stages of Drosophila melanogaster, from larvae to adult.
