Preparation method and application of polyclonal antibody against CCR12 of Epinephelus coioides

Abstract

The invention provides a preparation method and application of a polyclonal antibody against CCR12 of Epinephelus coioides. Currently, three XCR1-like chemokine receptors are discovered in fish, i.e., XCR1, XCR1L and CCR12, wherein XCR1L and CCR12 are only discovered in the genomes of fish, but little research is conducted on the functions of XCR1L and CCR12; and so far, in-vitro expression of CCR12 from fish has not been reported, no one has prepared a polyclonal antibody against CCR12 of Epinephelus coioides, and application of the polyclonal antibody against CCR12 has not been reported, either. To further reveal the migration or propagation of Epinephelus coioides CCR12 positive cells in tissue with parasitic infections and to identify the types of the CCR12 positive cells, a recombinant protein of the N-terminal region and three extracellular regions the CCR12 is expressed via a prokaryotic system and purified, and a rabbit anti-CCR12 recombinant protein polyclonal antibody is eventually prepared for the first time; and the antibody can specific recognize the CCR12 protein of Epinephelus coioides and mark cells in blood, so a foundation is laid for further marking of the CCR12 positive cells and research on parasitic infection resistance of the CCR12 positive cells.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a preparation method and application of a polyclonal antibody of CCR12 of grouper. Background technique [0002] Chemokines are a class of cytokines with very small molecular weight, and they have similar monomer structures (Baggiolini, 1998; Jin et al., 2005). According to their structure and the sequence of four conserved cysteines, chemokines can be divided into five types: CXC type, CC type, XC type, CX3C type and CX type (Murphy, et al., 2000; Nomiyama et al., 2008). To function, chemokines need to bind to their corresponding receptors. XCL1 is an XC-type chemokine that is produced by thymic myeloid epithelial cells, or activated T cells, NK cells, and NKT cells during homeostasis, pathogenic infection, or inflammatory response (Lei et al., 2012). XCR1 is the sole receptor for XCL1, mediated by CD8α + and CD11b - CD103 + Secreted by DC cells (Alexand...

AI Technical Summary

Problems solved by technology

Among them, XCR1L and CCR12 are only found in the fish genome, but there are few studies on their functions, and the lack of anti-XCR1L or CCR12 antibodies also limits the isolation and function of their positive cells.

So far, there is no report on the expression of fish CCR12 in vitro, let alone the research on CCR12 polyclonal antibody

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