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J. Biochem, 1988, Vol. 103, No. 2 225-230
© 1988 Japanese Biochemical Society

research-article

Monoclonal Antibodies against Rat T-Kininogen: Application to Radio immunoassay and Immunohistochemistry1

Iku Utsunomiya*, Sachiko Oh-ishi*,2, Izumi Hayashi*, Junji Maruhashi*, Naoki Tsuji*, Noboru Yamamoto** and Shohei Yamashina***

* Department of Pharmacology, School of Pharmaceutical Sciences, Kitasato University Minato-ku, Tokyo 108
** Department of Ultrastructural Research, School of Nursing, Kitasato University Sagamihara, Kanagawa 228
*** Department of Anatomy, School of Medicine, Kitasato University Sagamihara, Kanagawa 228

2To whom correspondence should be addressed.

Monoclonal antibodies to rat T-kininogen were produced and 9 hybridomas were selected. Radioimmunoassay (RIA) was developed using 125I-labeled T-kininogen and cell walls of Staphylococcus aureus (Zysorbin) for the separation of bound from free ligand, when IgG2a and IgG2b were used. In the case of IgG1 monoclonals, a second antibody (goat anti-mouse IgG) and Zysorbin were used. By this RIA, 1–16 ng T-kininogen/tube showed a linear inhibition curve, and cross reactivities to rat purified LMW- and HMW-kininogens were less than 0.5%, respectively. These monoclonal antibodies were also used for the immuno-histochemical staining of the liver to detect T-kininogen in hepatocytes. By using the RIA and immunohistochemical staining, the T-kininogen levels in rat plasma and liver following carrageenin-induced inflammation were estimated. At 3–5 h after the carrageenin injection, when the paw swelling was at its peak, the plasma level of T-kininogen and staining of the liver were slightly increased. T-Kininogen levels in plasma and liver peaked on the 2nd day, when the paw swelling had already decreased. The result indicates that the increase of T-kininogen level in the liver and plasma occurs with a time lag and T-kininogen is not directly involved in the increase of vascular permeability in carrageenin paw edema.

1This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.


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