Technoclone Anti-tPA Monoclonal Antibody 3VPA

IgG₁; half maximal binding to coated t-PA at 8.5 µg/mL (high affinity); Reaction with an epitope expressed on both the finger domain and growth factor domain of t-PA. Competitive inhibition of plasminogen activation in the presence of fibrin (Ki=0.68 nM). It can be used in an ELISA-system as second antibody.

Specificity

Binds to tPA. Reaction with an epitope expressed on both the finger domain and growth factor domain of tPA (1). Half maximal binding to coated t-PA AT 8.5 µg/ml (high affinity). Competitive inhibition of plasminogen activation in the presence of fibrin (Ki = 0.68nM).

Derivation

Hybridization of NSO cells with spleen cells from a BALB/c mouse immunized with purified melanoma-type t-PA.

Antibody class

IgG₁

Purification

The antibody is purified from murine ascites by affinity chromatography on immobilized recombinant Protein A. The purity of the product is tested by Laemmli SDS-PAGE.

Handling and storage

The antibody is lyophilized from a 1 mg/ml solution in isotonic phosphatebuffered saline, pH 7.4, containing 0.02% sodium azide and 20 mg/ml mannitol. It is supplied in vials of 500 µg and should be reconstituted with 0.5 ml distilled water. For extensive dilutions a protein containing solution should be used (e.g. 1% bovine serum albumin in PBS). Lyophilzed antibody should be stored at 4°C. Reconstituted antibody should be aliquoted and stored at -20°C or lower. Avoid repeated freeze-thaw cycles.

Literature

1. J.Wojta, L.Turcu, O.F.Wagner, C.Korninger, B.R.Binder: Evaluation of fibrinolytic capacity by a combined assay system for tissue-type plasminogen activator antigen and function using monoclonal anti-tissue-type plasminogen activator antibodies. J.Lab.Clin Med. 105:665 . 671, 1987. 2) J.Wojta, R.L.Hoover, and T.O.Daniel: Vascular Origin Determines Plasminogen Activator Expression in Human Endothelial Cells. J.Biol.Chem. 264 (5): 2846, 1989.