Immunogens used to prepare lrrc52 polyclonal antibodies
A technology of polyclonal antibodies and immunogens, applied in the direction of anti-animal/human immunoglobulins, antibodies, immunoglobulins from serum, etc., can solve the problems of contraceptive drugs that do not have male reproductive physiology
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Embodiment 1
[0029] Example 1 Design and preparation of LRRC52 polyclonal antibody (LID1)
[0030] 1. Prepared by Beijing Xinnuo Jinke Biotechnology Co., Ltd., the antibody is named LID1, and the preparation steps are as follows:
[0031] Such as Figure 13As shown (LRRC52 N-terminal 1-244 is extracellular, 245-265 is the transmembrane region, and human C-terminal 266-313 (mouse is 266-314) is located in the cytoplasm. The 84-98 polypeptide segment marked in the figure is used as the preparation of rabbit The antigenic site of the polyclonal antibody. The arrows correspond to the specific amino acid sequence of the polypeptide in humans and mice and the sequence comparison results, showing that humans and mice are completely homologous;
[0032] Analysis and design of peptide sequences: use DNAstar software to analyze the epitope of amino acids of LRRC52, evaluate its hydrophilicity, antigenicity, surface possibility, flexible region and other indexes, comprehensively consider the amino a...
Embodiment 2
[0036] 1. The LID1 antibody was prepared as in Example 1.
[0037] 2. Experimental animals and handling
[0038] In this experiment, 14-18 weeks old C57 male mice were purchased from the Model Animal Center of Nanjing University. The mice were free to eat and drink, the room temperature was controlled at 20-25 °C, and the lighting was given according to the law of 12 hours each day and night. After the male mice were killed by cervical dislocation, the abdominal cavity was exposed, and the bilateral epididymis were taken out. The fat and mesangial tissue were separated as much as possible under the dissecting microscope, washed 2 to 3 times, the tail was cut off, placed in a petri dish filled with HS, scratched, and placed at 37 °C, 5% CO 2 Incubate in an incubator for 20 min, centrifuge at 1600 rpm for 5 min to precipitate sperm, and wash twice with HS.
[0039] 3. Experimental method
[0040] 1. Separation of human sperm
[0041] Use Percoll discontinuous gradient centr...
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