2021PubMedRequires access

[Preparation and identification of mouse anti-Helicoverpa armigera sulfatase (HaSulf1) polyclonal antibody].

Xue Li, Kuerban Guli, Long Zhang, Xiaoli Ma

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Abstract

Objective To obtain the prokaryotic expression of Helicoverpa armigera sulfatase 1 (HaSulf1) protein and prepare its polyclonal antibodies. Methods A prokaryotic expression vector was firstly constructed. After inducing the protein expression, the crude recombinant protein was collected and purified with a nickel column, which was injected in immunized mice. Mouse blood was collected for detecting the titers of polyclonal antibodies by ELISA and the immune specificity was detected by Western blotting. Results The recombinant plasmid was obtained and the fusion protein was positively expressed and purified. ELISA data showed that the polyclonal antibody titer was 1:819 200. Western blotting showed that the prepared antibodies could bind to both the recombinant protein expressed in vitro and natural sulfatase from cotton bollworms. Conclusion Prokaryotic expression of recombinant HaSulf1 protein was obtained and specific mouse polyclonal antibodies were prepared.

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What this paper is about

Objective To obtain the prokaryotic expression of Helicoverpa armigera sulfatase 1 (HaSulf1) protein and prepare its polyclonal antibodies. Methods A prokaryotic expression vector was firstly constructed. After inducing the protein expression, the crude recombinant protein was collected and purified with a nickel column, which was injected in immunized mice. Mouse blood was collected for detecting the titers of polyclonal antibodies by ELISA and the immune specificity was detected by Western blotting. Results The recombinant plasmid was obtained and the fusion protein was positively expressed and purified. ELISA data showed that the polyclonal antibody titer was 1:819 200. Western blotting showed that the prepared antibodies could bind to both the recombinant protein expressed in vitro and natural sulfatase from cotton bollworms. Conclusion Prokaryotic expression of recombinant HaSulf1 protein was obtained and specific mouse polyclonal antibodies were prepared.

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Available abstract

Objective To obtain the prokaryotic expression of Helicoverpa armigera sulfatase 1 (HaSulf1) protein and prepare its polyclonal antibodies. Methods A prokaryotic expression vector was firstly constructed. After inducing the protein expression, the crude recombinant protein was collected and purified with a nickel column, which was injected in immunized mice. Mouse blood was collected for detecting the titers of polyclonal antibodies by ELISA and the immune specificity was detected by Western blotting. Results The recombinant plasmid was obtained and the fusion protein was positively expressed and purified. ELISA data showed that the polyclonal antibody titer was 1:819 200. Western blotting showed that the prepared antibodies could bind to both the recombinant protein expressed in vitro and natural sulfatase from cotton bollworms. Conclusion Prokaryotic expression of recombinant HaSulf1 protein was obtained and specific mouse polyclonal antibodies were prepared.

Key concepts: Polyclonal antibodies, Helicoverpa armigera, Recombinant DNA, Molecular biology, Blot, Antibody, Affinity chromatography, Titer

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[Preparation and identification of mouse anti-Helicoverpa armigera sulfatase (HaSulf1) polyclonal antibody]. — Research Paper | ScholarLens