1. Rat keap1 target information.
Rat Kelch-like ech-associated protein 1 (KEAP1) is a protein widely expressed in a variety of tissues, especially in metabolically active tissues such as liver, kidney, and intestine. Keap1 protein has multiple domains, including branched-chain-aa residue binding domain, broad complex, tramtrack, bric-a-brac (btb) domain, and kelch domain. These domains play an important role in the functioning of keap1.
As an important receptor protein, KEAP1 can recognize various stressors, such as oxidative stress and reactive nitrogen clusters, and interact with them, resulting in the activation of KEAP1-NRF2 signaling pathway. This process plays an important role in cellular oxidative stress defense and adaptation mechanisms. Studies have shown that the abnormality of KEAP1 can lead to the imbalance of redox balance in cells, which in turn leads to a series of diseases, such as cancer and neurodegenerative diseases.
2. ELISA method.
An enzyme-linked immunosorbent assay (ELISA) is a commonly used immunological assay to detect the presence of a specific antigen or antibody in a sample. The ELISA method has the advantages of high sensitivity, strong specificity, and easy operation, and is widely used in biomedical research, clinical diagnosis and other fields.
For the detection of KAP1 in rats, an indirect method can be used for ELISA assays. The specific steps are as follows:
1.The antigen is coated in the wells of the plate to form an antigen-antibody conjugate
2.Enzyme-labeled antibodies are added, which bind to antigen-antibody complexes;
3.The substrate is added for color development, and the substrate is made to produce colored products through an enzyme-catalyzed reaction
4.Stop the reaction so that the color change stops;
5.The optical density value of each well was measured with a microplate reader, and the concentration of the target protein in the sample to be measured was calculated according to the standard curve.
The detection of the expression level of KEAP1 in rats by ELISA method can provide reliable experimental data for related research, help to understand the mechanism of action of KEAP1 in the occurrence and development of diseases, and provide new ideas and methods for disease prevention and development. At the same time, the study of KEAP1 is also helpful to reveal the regulatory mechanism of cellular stress response and redox balance, and provides important theoretical basis and technical support for other related fields.