Materials and Reagents

Mice (C57BL/6, BALB/C)
Ascites of the trinitrophenyl (TNP)-specific IgE mAb from the IGELb4 (ATCC, catalog number: TIB141 )
TNP11-OVA (BioTechniques, catalog number: T-5051-10 )
Ovalbumin (OVA) (grade V) (Sigma Aldrich, catalog number: A5503 )
Phosphate buffered saline (PBS) (Life Technologies, Gibco^®, catalog number: 20012 )

Equipment

Needle (29G1/2)
Syringe (1 ml) (Terumo Medical Corporation, catalog number: SS-10M2913 )
Dial thickness guage caliper (Ozaki, catalog number: G-1A )

Procedure

Mice are sensitized with TNP-IgE mAb intravenous injection of ascites (200 μl).
One day later, the left ears of mice are injected subcutaneously with 10 μg of TNP11-OVA in 10 μl of PBS.
And their right ears are injected with equal amount of OVA as a control.
Ear thickness is measured with a dial thickness guage caliper for 7 days.

Acknowledgments

This protocol was developed and implemented by Dr. Masato Kubo at Division of Molecular Pathology, Tokyo University of Science, Chiba, Japan and Dr. Hiromasa Inoue at Department of Pulmonary Medicine, Kagoshima University, Kagoshima, Japan. This work was supported by a Grant-in-Aid-of-Scientific Research in Priority Areas of the Ministry of Education, Culture, Sports, Science, and Technology of Japan and the Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation.

References

Mukai, K., Matsuoka, K., Taya, C., Suzuki, H., Yokozeki, H., Nishioka, K., Hirokawa, K., Etori, M., Yamashita, M., Kubota, T., Minegishi, Y., Yonekawa, H. and Karasuyama, H. (2005). Basophils play a critical role in the development of IgE-mediated chronic allergic inflammation independently of T cells and mast cells. Immunity 23(2): 191-202.
Sawaguchi, M., Tanaka, S., Nakatani, Y., Harada, Y., Mukai, K., Matsunaga, Y., Ishiwata, K., Oboki, K., Kambayashi, T., Watanabe, N., Karasuyama, H., Nakae, S., Inoue, H. and Kubo, M. (2012). Role of mast cells and basophils in IgE responses and in allergic airway hyperresponsiveness. J Immunol 188(4): 1809-1818.