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Preparation and Application of Polyclonal Antibodies against GST

Meng Xu

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Abstract

Glutathione S transferase(GST) gene fusion system has being widely used in genetic engineering for expression of foreign proteins. Expressed fusion proteins can be easily detected and purified from bacterial lysates by antibodies against GST. pGEX plasmid vectors which contain GST gene are transformed to E.coli (TG1) for expression of GST in LB medium by inducing of 0.5mmol/L IPTG. The expressed GST was used to immunize rabbits after purification. The title of antiserum against GST after the third immunization was found more than 2×10 5. The preparation of antibodies has been successfully used in Western blot analysis and immunoaffinity chromatography of two novel GST fusion proteins(GST FLP,GST PreS1).

About this research paper

What this paper is about

Glutathione S transferase(GST) gene fusion system has being widely used in genetic engineering for expression of foreign proteins. Expressed fusion proteins can be easily detected and purified from bacterial lysates by antibodies against GST. pGEX plasmid vectors which contain GST gene are transformed to E.coli (TG1) for expression of GST in LB medium by inducing of 0.5mmol/L IPTG. The expressed GST was used to immunize rabbits after purification. The title of antiserum against GST after the third immunization was found more than 2×10 5. The preparation of antibodies has been successfully used in Western blot analysis and immunoaffinity chromatography of two novel GST fusion proteins(GST FLP,GST PreS1).

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Available abstract

Glutathione S transferase(GST) gene fusion system has being widely used in genetic engineering for expression of foreign proteins. Expressed fusion proteins can be easily detected and purified from bacterial lysates by antibodies against GST. pGEX plasmid vectors which contain GST gene are transformed to E.coli (TG1) for expression of GST in LB medium by inducing of 0.5mmol/L IPTG. The expressed GST was used to immunize rabbits after purification. The title of antiserum against GST after the third immunization was found more than 2×10 5. The preparation of antibodies has been successfully used in Western blot analysis and immunoaffinity chromatography of two novel GST fusion proteins(GST FLP,GST PreS1).

Key concepts: Polyclonal antibodies, Fusion protein, Antiserum, Molecular biology, lac operon, Affinity chromatography, Glutathione S-transferase, Western blot

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