Among the various immunization methods and injection sites available, Chondrex, Inc. highly recommends using subcutaneous administration at the base of the tail for immunizing mice. This method ensures consistent and effective exposure to an antigen, which elicits a robust antibody response. An important benefit of this technique is that it avoids altering or damaging the skin on the abdomen or back, which researchers may need for later experimental procedures and evaluations.
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Protocol
A. Materials
1. A Hamilton Syringe; A glass syringe must be used since plastic syringes generate high resistance for dispensing emulsions.
2. Needles (25 or 17-gause x 5/8")
3. A restraining device
B. Procedures
1. Place a 25 or 27-gauge x 5/8” needle on the Hamilton syringe. Before each injection, wipe the needle to prevent leakage of the emulsion.
2. Insert the needle bevel side up and parallel to the tail 2 cm from the base of the tail until the needle tip is 0.5 cm from the base. The entire needle should be subcutaneous.
Note: Insert the needle between the tail veins to avoid accidental injection into the bloodstream.
3. Inject 0.1 ml (100 µg antigen/mouse) of the emulsion subcutaneously at the base of the tail.
Note: If you need a booster injection, insert the needle at 3 cm from the base of the tail until the tip reaches 1.5 cm from the base. The booster injection should be administered at a different location from the initial injection.
