Ⅰ. Antigen:

protein, polypeptide, organelle, cell, tissue, etc.

Ⅱ. Immunization Mode:

Subcutaneous injection, intraperitoneal injection, intravenous injection, intramuscular injection, etc.

Ⅲ. The amount of antigens required for different animal immunity(take protein immunity as an example):

Subcutaneous injection, intraperitoneal injection, intravenous injection, intramuscular injection, etc.

	18-20kDa		<18-20kDa
Animal	Antigen quantity	Optimal antigen quantity	Antigen quantity	Optimal antigen quantity
Rabbit	20-200	100	50-400	200
Mice	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: μg

Ⅳ. Immunization Program (take immunized rabbit as an example):

Days	0	14	28	38	56	66	87
Injection	x	x	x		x
Collect blood	2ml			2ml		2+20ml	2+50-70ml

Ⅴ. Immunization Program (take immunized rabbit as an example):

In the first immunization, the complete adjuvant was mixed with immunogen, in the later, the incomplete adjuvant was mixed with immunogen, and multi-point, time interval immunization was used.

If the rabbit is immunized with cells, the amount of cells required for each immunization is 2-3 x 10⁷ cells.

After three times of continuous immunization, a small amount of blood needs to be taken for ELISA to detect the antibody titer of the immunized animals. Generally, the titer can reach 1:10000-50000. The immunized animals should be immunized once a week before being killed.

If immunized with mice, small amount of blood can be collected from tail vein (about 50 μ L), and large amount of blood can be collected from eyeball (about 500-1000 μ L); if immunized with rabbits, small amount of blood can be collected from ear edge vein (about 2ml), and large amount of blood can be collected from heart (50-100ml)

According to the standard method of serum preparation, the serum was separated and packed into small parts and stored at -80℃.

Related reading：

Western Blot

Indirect ELISA To Detect Specific Antibodies