Preparation and Detection of Polyclonal Antibody Against TLR2 Receptor in Cyprinus Carpio
Toll-like receptor 2(TLR2)is one of the important constituent members of the TLR family and plays an important role and significance in the congenital immune system.Studies have shown that the function of TLR2 in bony fish is more extensive than previously predicted and plays an important role in the anti-bacterial immunity of the host.In order to obtain the polyclonal antibody of carp TLR2 and detect it.In this experiment,PCR technique was used to amplify the TLR2 gene from the TLR2 receptor antigen in the Carp region,and it was cloned to the expression vector pET-32a(+)to construct recombinant prokaryotic expression plasmid,and the ex-pression was induced in Escherichia coli,and then the antigen was purified by His column.The TLR2 polyclonal antibody was obtained by using the purified antigen immune rabbits.Immunohistochemistry was used to detect the specificity of TLR2 polyclonal antibodies.The results showed that the recombinant expression plasmid of pet-32a(+)-TLR2 was successfully constructed and expressed in Escherichia coli,and the purified recombinant protein was obtained,the molecular weight was about 41.8 ku,the antibody titer was 3×104,and the polyclonal antibody could specifically identify the TLR2 receptor protein in the intestinal tissue of carp.The results showed that the prepared antibodies had high potency,affinity and specificity.Successful preparation of multi-clone antibodies a-gainst carp TLR2 receptors in rabbits to further study the relationship between the expression of TLR2 protein and the health of carp and the role played in immune response lay the foundation.
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