Monoclonal antibodies against rat T-kininogen: application to radioimmunoassay and immunohistochemistry.
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 immunohistochemical 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.
Utsunomiya
I
I
Department of Pharmacology, School of Pharmaceutical Sciences, Kitasato University, Tokyo.
Oh-ishi
S
S
Hayashi
I
I
Maruhashi
J
J
Tsuji
N
N
Yamamoto
N
N
Yamashina
S
S
D000818:Animals; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay; D007114:Immunization; D007150:Immunohistochemistry; D007704:Kininogens / Q000032:analysis*; D011863:Radioimmunoassay; D051381:Rats
D000911:Antibodies, Monoclonal; D007704:Kininogens