Tissue type Plasminogen Activator Activity Assay Kit (Colorimetric, Human) (ab108905)
Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Cell culture supernatant, Plasma
- Sensitivity: 0.03 IU/ml
Overview
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Product name
Tissue type Plasminogen Activator Activity Assay Kit (Colorimetric, Human)
See all Tissue Plasminogen Activator kits -
Detection method
Colorimetric -
Sample type
Cell culture supernatant, Plasma -
Assay type
Enzyme activity (quantitative) -
Sensitivity
0.03 IU/ml -
Range
0.039 IU/ml - 40 IU/ml -
Assay time
1h 00m -
Species reactivity
Reacts with: Human -
Product overview
Tissue type Plasminogen Activator Activity Assay Kit (Colorimetric, Human) ab108905 is used to measure Human Tissue type Plasminogen Activator activity in cell culture supernatants and plasma.
The tPA assay protocol measures the ability of Tissue type Plasminogen Activator to activate the plasminogen to plasmin in coupled or indirect assays that contain Tissue type Plasminogen Activator, plasminogen, and a plasmin-specific synthetic substrate. The amount of plasmin produced is quantitated using a highly specific plasmin substrate releasing a yellow para-nitroaniline (pNA) chromophore. The change in absorbance of the pNA in the reaction solution at 405 nm is directly proportional to the Tissue type Plasminogen Activator enzymatic activity.
The entire kit may be stored at -20°C for long term storage before reconstitution - Avoid repeated freeze-thaw cycles.
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Notes
Previously called Tissue type Plasminogen Activator Human Chromogenic Activity Assay Kit.
Tissue type Plasminogen Activator (tPA) is a 68 kDa serine protease that converts the zymogen plasminogen into the active serine protease plasmin which digests fibrin and induces the dissolution of fibrin clots. Tissue type Plasminogen Activator is synthesized by endothelial cells in normal blood vessels and displays relatively high affinity for fibrin, suggesting that it functions predominately in physiological thrombolysis in vivo.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests 1X Diluent 1 x 30ml Human Plasminogen 1 unit Microplate 1 96 well polystyrene microplate (12 strips of 8 wells) 1 unit Plasmin Substrate 2 units Sealing Tapes 3 units Tissue type Plasminogen Activator Standard 1 x 32 units -
Research areas
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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. Play a direct role in facilitating neuronal migration. -
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. -
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. -
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. -
Domain
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. -
Post-translational
modificationsThe 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. -
Cellular localization
Secreted > extracellular space. - Information by UniProt
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Alternative names
- Alteplase
- DKFZp686I03148
- Plasminogen activator tissue
see all -
Database links
- Entrez Gene: 5327 Human
- Omim: 173370 Human
- SwissProt: P00750 Human
- Unigene: 491582 Human
Associated products
Images
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Functional Studies- Tissue Type Plasminogen Activator Human ELISA Kit (ab108905)
tPA measured in biological fluids showing activity (IU) per mL of tested sample
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Functional Studies- Tissue Type Plasminogen Activator Human ELISA Kit (ab108905)Time course of tPA measured in biological fluids showing optical densities
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Functional Studies- Tissue Type Plasminogen Activator Human ELISA Kit (ab108905)Standard curve (colorimetric) : mean of duplicates (+/-SD) with background readings subtracted
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Functional Studies- Tissue Type Plasminogen Activator Human ELISA Kit (ab108905)Change of absorbance per min. of various concentrations of tPA standard
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Functional Studies- Tissue Type Plasminogen Activator Human ELISA Kit (ab108905)O.D's of various dilutions of human plasma after 20hrs using ab108905
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Functional Studies - Tissue type Plasminogen Activator Human ELISA Kit (ab108905)Tissue type Plasminogen Activator Chromogenic Activity Standard curve
References (3)
ab108905 has been referenced in 3 publications.
- Faine BA et al. Helicopter "Drip and Ship" Flights Do Not Alter the Pharmacological Integrity of rtPA. J Stroke Cerebrovasc Dis 27:2720-2724 (2018). PubMed: 30037651
- Nabiabad HS et al. Expression of active chimeric-tissue plasminogen activator in tobacco hairy roots, identification of a DNA aptamer and purification by aptamer functionalized-MWCNTs chromatography. Protein Expr Purif 152:137-145 (2018). PubMed: 26876003
- Gupta V et al. Glaucoma is associated with plasmin proteolytic activation mediated through oxidative inactivation of neuroserpin. Sci Rep 7:8412 (2017). PubMed: 28827627
