[Expression and purification of p11 fusion protein in E. coli and preparation of antiserum against GST-p11].
OBJECTIVE
To obtain p11 fusion protein and prepare specific polyclonal antibody against p11.


METHODS
A full-length human p11 gene was cloned into expression vectors, pGEX-4T-2 and pQE30, and transformed into E. coli. The expressed proteins were purified from lysates with Glutathione Sepharose 4B and the Ni-NTA agarose column, respectively. The purified GST-p11 was mixed with Freund's complete or incomplete adjuvant and immunized rabbits.


RESULTS
A high level of expression of target proteins was detected after IPTG induction and purified proteins were obtained by affinity chromatography with Glutathione Sepharose 4B and the Ni-NTA agarose column, respectively. Western blotting analysis suggested that the polyclonal antibody can recognize 6xHis-p11 and GST protein.


CONCLUSIONS
The antiserum against p11 prepared by prokaryotic expression of GST-p11 fusion protein has good specificity.
Tan
Zhi-ping
ZP
National Laboratory of Medical Genetics, Central South University, Changsha 410078, China.
Huang
Liang-qun
LQ
Zheng
Duo
D
D000818:Animals; D017306:Annexin A2; D000906:Antibodies / Q000276:immunology; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology*; D008297:Male; D011387:Prokaryotic Cells / Q000378:metabolism; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D009418:S100 Proteins; D000099248:S100 Calcium Binding Protein A10
D017306:Annexin A2; D000906:Antibodies; D002135:Calcium-Binding Proteins; D005982:Glutathione Transferase; D007106:Immune Sera; D011993:Recombinant Fusion Proteins; D009418:S100 Proteins; D000099248:S100 Calcium Binding Protein A10