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Laboratory animal immunization protocol
1. Antigens: proteins, peptides, organelles, cells, tissues, etc.
2. Immunization method: subcutaneous injection, intraperitoneal injection, intravenous injection, intramuscular injection, etc.
3. The amount of antigens required for different animal immunizations (taking protein immunization as an example)
> 18-20kDa <18-20kDa
Animal antigen amount optimal antigen amount antigen amount optimal antigen amount
Rabbit 20-200 100 50-400 200
Mouse 2-40 15 4-60 40
Rat 10-50 30 20-150 50
Sheep 100-1000 200 200-1000 400
Hen 20-200 100 50-400 200
Unit: microgram
4. Immunization plan (take immunization rabbit as an example):
Days 0 14 28 38 56 66 87
Injection x x x x
Blood collection 2ml 2ml 2 + 20ml 2 + 50-70ml
5. Note:
1) The complete adjuvant is mixed with the immunogen for the first immunization, and the incomplete adjuvant is mixed with the immunogen for the booster immunization. A multi-point, time-interval immunization method is adopted.
2) If the rabbit is immunized with cells, the amount of cells required for each immunization is 2-3 x 107 cells.
3) After three consecutive immunizations, a small amount of blood is required for ELISA detection to detect the antibody titer of the immunized animal, generally the titer can reach 1: 10,000-50,000. Immunization should be boosted once a week before the animals immunized.
4) If you are immunized with mice, you can collect a small amount of blood from the tail vein (about 50µl), and finally take a large amount of blood from the eyeball (about 500-1000µl); if you are immunized with a rabbit, you can take a small amount of blood from the ear vein (about 2mL), and finally Large blood collection of the heart (50-100mL)
5) Separate the serum according to the standard method of serum preparation, divide it into small portions, and store at -80oC.
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