Method for preparing antibody aiming at beet armyworm caspase-1

Abstract

The invention relates to a method for preparing antibody aiming at beet armyworm caspase-1. The method comprises the following steps: immunizing an animal through polypeptide as shown in SEQ ID NO: 1 of an amino acid sequence, collecting serum of the animal, and purifying antibody from the serum. The invention further discloses the antibody obtained through the method. The invention further discloses a method for detecting beet armyworm caspase-1 large-subunit through the antibody. Through the adoption of the method and the antibody, the beet armyworm caspase-1 large-subunit in a sample can be specifically detected, and moreover, excitation of caspase-1 protein during insect cell apoptosis is initially detected, that is, breakage of large and small-subunit is detected. The method has important significance for the research on the toxicological meaning of the insect caspase gene family and the development of a new pesticide taking caspase as a target.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a method for preparing antibodies against the large subunit of caspase-1 of beet armyworm. Background technique [0002] Caspases are a class of conserved aspartate-specific cysteine ​​proteases present in multicellular organisms. As the central effector of apoptosis, caspase plays a key role in the initiation and process of apoptosis. Members of the caspase family complete the cascade reaction among caspase members through the recruitment, transmission and execution of apoptotic signals to promote the process of cell apoptosis. The caspase family is divided into two subfamilies of inflammatory and apoptotic caspases according to their physiological functions. Apoptotic caspases can be divided into initiator caspases and effector caspases. Upon initiation of apoptosis, activation of initiating caspases is a critical point at which cells choose between apoptosis or survival, ma...

AI Technical Summary

Problems solved by technology

The inventor once tried to select the full-length caspase-1 as the immunogen to make antibodies, but found that the rabbits could not tolerate it during the process of immunizing rabbits, and the inventors repeatedly selected some peptide segments as antigens as antibodies, Antibody not needed

For example, see Figure 4 , the inventor selected part of the N-terminal prodomain and the entire large subunit sequence (29-195aa) as an immunogen to immunize rabbits, and the resulting serum could only detect the caspase-1 holoenzyme, but the caspase-1 large subunit could not be detected

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