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Updated: Apr 5, 2026
A High Content Imaging Assay for Identification of Botulinum Neurotoxin Inhibitors
Published on: November 14, 2014
[Preparation and application of polyclonal antibody against Ras-related C3 botulinum toxin substrate 1]
Lili Zhang1, Lu Hao2, Wei Zuo2
1School of Graduate, Anhui Medical University, Hefei 230032; Department of Antibody Engineering, Beijing Proteome Research Center, State Key Laboratory of Proteomics, Beijing Institute of Radiation Medicine, Academy of Military Medicine, Beijing 102206, China.
Objective:
To prepare rabbit polyclonal antibody (pAb) against Ras-related C3 botulinum toxin substrate 1 (Rac1) and identify the functions of the antibody.
Methods:
The Rac1 fragment was amplified from cDNA of human cervical cancer HeLa cells, and then inserted into the vector pET-32a to construct the recombinant plasmid pET-32a-Rac1. The recombinant protein Rac1 was fused with His-tag and the fusion protein His-Rac1 was expressed in E.coli expression system. Thereafter, His-tagged Rac1 was purified to immunize rabbits as immunogen. The titer and specificity of rabbit polyclonal antibody against Rac1 were identified by ELISA, Western blotting and immunohistochemistry, respectively.
Results:
The fusion protein His-Rac1 was successfully expressed in E.coli. The pAb could specifically detect Rac1 in human HeLa cells and MCF-7 cells in Western blotting. Moreover, the pAb could specifically recognized Rac1 protein in human kidney tissues.
Conclusion:
The pAb of human Rac1 protein was successfully prepared, which could specifically recognize Rac1 protein in several natural samples.
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