|Table of Contents|

Preparation of polyclonal antibody against human KIAA0100 protein

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2017 05
Page:
681-686
Research Field:
Publishing date:

Info

Title:
Preparation of polyclonal antibody against human KIAA0100 protein
Author(s):
Cui HeZhang Wanggang
Department of Clinical Hematology,Affiliated No.2 Hospital,Xi'an Jiaotong University,Shaanxi Xi'an 710004,China.
Keywords:
human KIAA0100 proteinpAbprokaryotic expressionrecombinant plasmidrecombinant protein
PACS:
R730.51
DOI:
10.3969/j.issn.1672-4992.2017.05.003
Abstract:
Objective:To preparation of polyclonal antibody(pAb) against human KIAA0100 protein and lay the foundation for the study of the structure and function of human KIAA0100 protein.Methods:Bioinformatics software was used to optimize Escherichia coli(E.coli) rare codons into E.coli normal codons of human KIAA0100 protein segment(1 557~2 234),and then the synthetic optimal nucleotide sequence were cloned into prokaryotic expression vector pET-30a to construct the recombinant plasmid.After being identified by restriction enzyme digestion analysis,the recombinant plasmid were transformed into E.coli BL21(DE3) cells,and then the recombinant human KIAA0100 protein segment(1 557~2 234) were expressed as a fusion protein with C-terminal 6×histidine(His)-tag.The New Zealand white rabbits were next immunized by the recombinant protein.The pAb was finally identificated by Western blot analysis.Results:Restriction enzyme digestion analysis showed that the recombinant plasmid was successfully constructed.SDS-PAGE analysis attested that the recombinant protein was successfully expressed as inclusion body.Western blot analysis proved that the pAb could sensitively identified human KIAA0100 protein.Conclusion:In this study,the pAb against human KIAA0100 protein is successfully prepared,which will be of great help on the study of the structure and function of human KIAA0100 protein.

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Memo:
国家自然科学基金资助项目(编号:0817 18110232)
Last Update: 2017-01-26