Laboratory animal immunization protocol

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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