The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent dilution.
1/50 - 1/100.
1/50. Pretreatment with 0.05% Pronase E for 20 minutes is recommended.
Converts the abundant, but inactive, zymogen plasminogen to plasmin by hydrolyzing a single Arg-Val bond in plasminogen. By controlling plasmin-mediated proteolysis, it plays an important role in tissue remodeling and degradation, in cell migration and many other physiopathological events. Play a direct role in facilitating neuronal migration.
Synthesized in numerous tissues (including tumors) and secreted into most extracellular body fluids, such as plasma, uterine fluid, saliva, gingival crevicular fluid, tears, seminal fluid, and milk.
Involvement in disease
Note=Increased activity of TPA results in increased fibrinolysis of fibrin blood clots that is associated with excessive bleeding. Defective release of TPA results in hypofibrinolysis that can lead to thrombosis or embolism.
Both FN1 and one of the kringle domains are required for binding to fibrin. Both FN1 and EGF-like domains are important for binding to LRP1. The FN1 domain mediates binding to annexin A2. The second kringle domain is implicated in binding to cytokeratin-8 and to the endothelial cell surface binding site.
The single chain, almost fully active enzyme, can be further processed into a two-chain fully active form by a cleavage after Arg-310 catalyzed by plasmin, tissue kallikrein or factor Xa. Differential cell-specific N-linked glycosylation gives rise to two glycoforms, type I (glycosylated at Asn-219) and type II (not glycosylated at Asn-219). The single chain type I glycoform is less readily converted into the two-chain form by plasmin, and the two-chain type I glycoform has a lower activity than the two-chain type II glycoform in the presence of fibrin. N-glycosylation of Asn-152; the bound oligomannosidic glycan is involved in the interaction with the mannose receptor. Characterization of O-linked glycan was studied in Bowes melanoma cell line.
Tissue-type plasminogen activator chain B antibody
Immunohistochemistry (Frozen sections) - Anti-TPA Tissue Plasminogen Activator antibody (ab14198)Image from Lohman RJ et al., Neurobiol Dis. 2008;30(1):84-93. Fig 2.; doi: 10.1016/j.nbd.2007.12.010 with permission from Elsevier.
a14198 staining TPA Tissue Plasminogen Activator (A - blue) in Rat hippocampal/thalamic region tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde. Tissue was incubated in pre-block solution (10% normal hourse serum in antibody diluent (PBS + 0.1% Trition X-100)) for 1 hour. Samples were incubated with primary antibody (1/50 in PBS + pre-block solution) for 16 hours. B - trypsin (red), C - PAR2 (green), D - merge.
TPA Tissue Plasminogen Actvator and Trypsin co-localise.
Immunohistochemistry (Frozen sections) - Anti-TPA Tissue Plasminogen Activator antibody (ab14198)This image is courtesy of an Abreview submitted by Mr Rink-Jan Lohman
ab14198 at 1/50 staining rat brain hippocampus pyramidal neuron tissue sections by IHC-Fr. The tissue was blocked with horse serum and incubated for 16 hours with the antibody. An AMCA conjugated donkey antibody was used as the secondary.
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