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Preparation and indentification of polyclonal antiserum against Em18 antigen

Chuntao Zhang

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Abstract

Objective: To obstain polyclonal antiserum against Em18 antigen for the foundation of further screening for Em18 related peptides. Methods: The recombinant expression plasmid pET41a-Em18 was transformed into BL21(E.coli) via IPTG induction and purified rEm18-GST as immunogen was used to immunize BALB/c mice. The specificity of mice anti-Em18-GST antibody was analyzed by Western blot and ELISA. Results: (1) The expressed rEm18-GST recombinant protein can be detected as a band of 50 kDa by SDS-PAGE.(2) High purity rEm18-GST recombinant protein was obtained after purification.(3) The polyclonal anti-Em18-GST antibody could specifically recognize rEm18-GST recombinant protein. ELISA showed a high titre (1∶25 600) of the antiserum. Conclusion: Polycloned anti-Em18-GST antibody with high specificity and purity has been prepared by using purified rEm18-GST as immunogen, which lays the foundation for further screening for Em18 related peptides.

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What this paper is about

Objective: To obstain polyclonal antiserum against Em18 antigen for the foundation of further screening for Em18 related peptides. Methods: The recombinant expression plasmid pET41a-Em18 was transformed into BL21(E.coli) via IPTG induction and purified rEm18-GST as immunogen was used to immunize BALB/c mice. The specificity of mice anti-Em18-GST antibody was analyzed by Western blot and ELISA. Results: (1) The expressed rEm18-GST recombinant protein can be detected as a band of 50 kDa by SDS-PAGE.(2) High purity rEm18-GST recombinant protein was obtained after purification.(3) The polyclonal anti-Em18-GST antibody could specifically recognize rEm18-GST recombinant protein. ELISA showed a high titre (1∶25 600) of the antiserum. Conclusion: Polycloned anti-Em18-GST antibody with high specificity and purity has been prepared by using purified rEm18-GST as immunogen, which lays the foundation for further screening for Em18 related peptides.

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Available abstract

Objective: To obstain polyclonal antiserum against Em18 antigen for the foundation of further screening for Em18 related peptides. Methods: The recombinant expression plasmid pET41a-Em18 was transformed into BL21(E.coli) via IPTG induction and purified rEm18-GST as immunogen was used to immunize BALB/c mice. The specificity of mice anti-Em18-GST antibody was analyzed by Western blot and ELISA. Results: (1) The expressed rEm18-GST recombinant protein can be detected as a band of 50 kDa by SDS-PAGE.(2) High purity rEm18-GST recombinant protein was obtained after purification.(3) The polyclonal anti-Em18-GST antibody could specifically recognize rEm18-GST recombinant protein. ELISA showed a high titre (1∶25 600) of the antiserum. Conclusion: Polycloned anti-Em18-GST antibody with high specificity and purity has been prepared by using purified rEm18-GST as immunogen, which lays the foundation for further screening for Em18 related peptides.

Key concepts: Immunogen, Polyclonal antibodies, Antiserum, Recombinant DNA, Molecular biology, Western blot, Antibody, Antigen

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