[Expression, purification, antibody preparation and identification of tumor protein translationally-controlled 1 (TPT1)].
OBJECTIVE
To express and purify the fusion protein of TPT1 (tumor protein translationally-controlled 1) in prokaryocytes and to prepare rabbit anti-TPT1 antibody.


METHODS
The expression vector pRSETA(2)-TPT1 was reconstructed and transformed into BL21 (DE3). TPT1 fusion protein was induced by IPTG and the TPT1 fusion protein purified by Ni-NTA His Bind resin was used to immunize the rabbit. The titer of the polyclonal antibody was detected by ELISA and its specificity was analyzed by Western blot and IF (immunofluorescence).


RESULTS
The TPT1 fusion protein was highly expressed in E.coli and specific polyclonal antibody was obtained after the immunization.


CONCLUSIONS
The recombinant prokaryotic expression vector pRSETA(2)-TPT1 is successfully constructed and high expression of TPT1 is induced in E.coli, which may lay the basis for further study of the development and treatment of tumors and other diseases.
Tianjin Life Science Research Center, Basic Medical School, Tianjin Medical University, Tianjin, China.
Yi-fan
YF
Zhao
Hai-fang
HF
Li
Shi-ming
SM
Tang
Meng
M
Wang
Hai-dong
HD
Wu
Min
M
Liu
Xin
X
Li
Hua
H
Tang
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification*; D014408:Biomarkers, Tumor / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D000090324:Tumor Protein, Translationally-Controlled 1
D000906:Antibodies; D014408:Biomarkers, Tumor; D011993:Recombinant Fusion Proteins; C000715647:TPT1 protein, human; D000090324:Tumor Protein, Translationally-Controlled 1