Sheep tPA - Tissue-type Plasminogen Activator Polyclonal Antibody | anti-PLAT antibody

Sheep anti-human tissue Plasminogen Activator (tPA), Affinity-purified IgG

Cat. Num. AAA512164

• Gene Names: PLAT; TPA; T-PA
• Purity: Affinity purified IgG.; Vial containing ml of IgG purified by affinitychromatography on immobilized tPA. Total protein is 0.5 mg.
• Synonyms: tPA - Tissue-type Plasminogen Activator; Polyclonal Antibody; Sheep anti-human tissue Plasminogen Activator (tPA); Affinity-purified IgG; human; anti-PLAT antibody

• Host: Sheep
• Clonality: Polyclonal
• Specificity: This antibody is specific for tPA as demonstrated by immunoelectrophoresis and ELISA.
• Purity/Purification: Affinity purified IgG.; Vial containing ml of IgG purified by affinitychromatography on immobilized tPA. Total protein is 0.5 mg.
• Form/Format: Affinity-purified IgG (APIgG), clear liquid.
• Concentration: APIgG concentration is mg/ml, determined by absorbance using an extinction coefficient (E1%280) of 13.4. (varies by lot)

• Application Notes: Suitable as a source of enriched antibodies to human tPA.
• Immunogen: Human tissue-type plasminogen activator prepared from melanoma cell line.
• Buffer: 10 mM HEPES, pH 7.4, 150 mM NaCl, 50% (v/v) glycerol.
• Neutralizing Activity: Not determined.
• Preparation and Storage: Store between -10 and -20 degree C. Product will become viscous but will not freeze. Avoid storage in frost-free freezers. Keep vial tightly capped. Allow product to warm to room temperature and gently mix before use.

Related Product Information for anti-PLAT antibody

Tissue-type plasminogen activator (tPA) is one of two major physiologic activators of plasminogen in plasma. It is a serine protease of 68 kDa produced primarily in endothelial cells but is also present in monocytes and megakaryocytes. Normal plasma tPA antigen concentrations have been reported from 20 ng/ml to 5 ug/ml, depending on the assay used, but typically most of the tPA (> 90%) is in complex with it's primary inhibitor, plasminogen activator inhibitor-1 (PAI-1). Structurally, tPA is a single-chain enzyme that consists of a catalytic domain followed by two kringle structures, an EGF domain and a finger domain. The activation of plasminogen by tPA is dependent on the presence of a fibrin cofactor. The binding of both tPA and plasminogen to fibrin is mediated in part through lysine binding sites within the kringle structures of both enzyme and substrate, but also through the finger domain of tPA. Activation of plasminogen by tPA occurs by cleavage after residue Arg560 to produce the two-chain active serine protease plasmin. The activity of tPA is regulated in part by a very short half life in circulation (t½ of ~4 minutes) and by circulating protease inhibitors PAI-1 and to a lesser extent alpha2macroglobulin1-3.

References

1. Bachmann F; The Plasminogen-Plasmin Enzyme System; in Hemostasis and Thrombosis, 3rd Edition, eds. RW Colman, J Hirsh, VJ Marder and EW Salzman, pp. 1592-1622, J.B. Lippincott Co., Philadelphia PA, USA, 1994.
2. Holvoet P, Cleemput H, Collen D; Assay of Human Tissue-Type Plasminogen Activator (t-PA) with an Enzyme-Linked Immunosorbent Assay (ELISA) Based on Three Murine Monoclonal Antibodies to t-PA. Thrombosis and Haemostasis 54, pp. 684-687, 1985.
3. Giles AR, Nesheim, et al; The fibrinolytic Potential of the Normal Primate following the Generation of Thrombin In Vivo; Thrombosis and Haemostasis 63, pp. 476-481, 1990.

NCBI and Uniprot Product Information

• NCBI GI #: 14702169
• NCBI GeneID: 5327
• NCBI Accession #: NP_127509.1
• NCBI GenBank Nucleotide #: NM_033011.2
• UniProt Accession #: P00750; Q15103; Q503B0; Q6PJA5; Q7Z7N2; Q86YK8; Q9BU99; Q9BZW1; A8K022; B2R8E8
• Molecular Weight: 62,917 Da
• NCBI Official Full Name: tissue-type plasminogen activator isoform 3 preproprotein
• NCBI Official Synonym Full Names: plasminogen activator, tissue
• NCBI Official Symbol: PLAT
• NCBI Official Synonym Symbols: TPA; T-PA
• NCBI Protein Information: tissue-type plasminogen activator; alteplase; reteplase; t-plasminogen activator; plasminogen activator, tissue type; tissue plasminogen activator (t-PA)
• UniProt Protein Name: Tissue-type plasminogen activator
• Protein Family: Tissue-type plasminogen activator
• UniProt Gene Name: PLAT
• UniProt Synonym Gene Names: t-PA; t-plasminogen activator; tPA
• UniProt Entry Name: TPA_HUMAN

NCBI Description

This gene encodes tissue-type plasminogen activator, a secreted serine protease which converts the proenzyme plasminogen to plasmin, a fibrinolytic enzyme. Tissue-type plasminogen activator is synthesized as a single chain which is cleaved by plasmin to a two chain disulfide linked protein. This enzyme plays a role in cell migration and tissue remodeling. Increased enzymatic activity causes hyperfibrinolysis, which manifests as excessive bleeding; decreased activity leads to hypofibrinolysis which can result in thrombosis or embolism. Alternative splicing of this gene results in multiple transcript variants encoding different isoforms. [provided by RefSeq, Jul 2008]

Uniprot Description

Function: 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. Plays a direct role in facilitating neuronal migration.

Catalytic activity: Specific cleavage of Arg-|-Val bond in plasminogen to form plasmin.

Enzyme regulation: Inhibited by SERPINA5. Ref.24

Subunit structure: Heterodimer of chain A and chain B held by a disulfide bond. Forms a heterodimer with SERPINA5. Binds to fibrin with high affinity. This interaction leads to an increase in the catalytic efficiency of the enzyme between 100-fold and 1000-fold, due to an increase in affinity for plasminogen. Similarly, binding to heparin increases the activation of plasminogen. Binds to annexin A2, cytokeratin-8, fibronectin and laminin. Binds to mannose receptor and the low-density lipoprotein receptor-related protein (LRP1); these proteins are involved in TPA clearance. Yet unidentified interactions on endothelial cells and vascular smooth muscle cells (VSMC) lead to a 100-fold stimulation of plasminogen activation. In addition, binding to VSMC reduces TPA inhibition by PAI-1 by 30-fold. Binds LRP1B; binding is followed by internalization and degradation. Ref.25

Subcellular location: Secreted › extracellular space.

Tissue specificity: 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.

Domain: Both FN1 and one of the kringle domains are required for binding to fibrin. Ref.23 Ref.33 Ref.34Both FN1 and EGF-like domains are important for binding to LRP1. Ref.23 Ref.33 Ref.34The FN1 domain mediates binding to annexin A2. Ref.23 Ref.33 Ref.34The second kringle domain is implicated in binding to cytokeratin-8 and to the endothelial cell surface binding site. Ref.23 Ref.33 Ref.34

Post-translational modification: 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. Ref.21 Ref.22N-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.

Involvement in disease: 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.

Pharmaceutical use: Available under the names Activase (Genentech) and Retavase (Centocor and Roche) [Retavase is a fragment of TPA that contains kringle 2 and the protease domain; it was also known as BM 06.022]. Used in Acute Myocardial Infarction (AMI), in Acute Ischemic Stroke (AIS) and Pulmonary Embolism (PE) to initiate fibrinolysis.

Sequence similarities: Belongs to the peptidase S1 family.Contains 1 EGF-like domain.Contains 1 fibronectin type-I domain.Contains 2 kringle domains.Contains 1 peptidase S1 domain.