Preparation and Application Analysis of a Polyclonal Antibody as Reference Protein in Helicoverpa armigera (Lepidoptera: Noctuidae)

Abstract

A stable and specific heat shock protein 27.2 (HSP27.2) antibody was prepared and analyzed for protein level research in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). The full-length hsp27.2 was amplified from H. armigera larvae and constructed into the prokaryotic expression vector. The purified His-tag fused protein was used to immunize rabbits for the antibody preparation. Western blot analysis indicated that this antibody specifically recognized the HSP27.2 encoded by H. armigera and detected the HSP27.2 encoded by other noctuid larvae. Further analysis of HSP27.2 expression in H. armigera under infection by different pathogenic microorganisms and in different tissues showed that the expression of HSP27.2 is continually stable. The HSP27.2 antibody is efficient and capable as a reference antibody for functional studies involving genes and proteins in H. armigera and other lepidopteran insects.