Overview

  • Product name

    Human Tissue Plasminogen Activator ELISA Kit
    See all Tissue type Plasminogen Activator kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Human serum 5 3.7%
    Inter-assay
    Sample n Mean SD CV%
    Human serum 3 10.5%
  • Sample type

    Cell culture supernatant, Serum, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    3.5 pg/ml
  • Range

    78 pg/ml - 5000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 104 100% - 107%
    Cell culture media 119 116% - 123%
    Hep Plasma 97 94% - 99%
    EDTA Plasma 84 82% - 86%
    Cit plasma 99 97% - 107%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Goat, Cow, Pig
  • Product overview

    Tissue Plasminogen Activator in vitro SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Tissue Plasminogen Activator (tPA) protein in human serum, plasma and cell culture supernatant samples.


    The SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Tissue Plasminogen Activator (Tissue-type plasminogen activator or tPA) is a circulating serine protease involved in the breakdown of clots. tPA converts inactive plasminogen to active plasmin; in turn plasmin degrades the fibrin matrix in clots. In addition, plasmin can cleave tPA at Arg-310 with results in a two chain disulphide linked tPA that has even greater proteolytic activity. tPA is synthesized in many tissues and is secreted into most extracellular body fluids. Recombinant tPA is used medically to resolve or prevent blood clots in ischemic stroke or myocardial infarction.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human TPA Capture Antibody 1 x 600µl
    10X Human TPA Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BI 1 x 6ml
    Human TPA Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • 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
    modifications

    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.
  • Cellular localization

    Secreted > extracellular space.
  • Information by UniProt
  • Alternative names

    • Alteplase
    • Plasminogen activator tissue
    • Plasminogen activator tissue type
    • PLAT
    • Reteplase
    • t-PA
    • T-plasminogen activator
    • Tissue-type plasminogen activator chain B
    • tPA
    • TPA_HUMAN
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab190812 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD, n =2) are graphed.

  • PBMC were grown in the absence or presence of phytohemagglutinin (PHA) for 2 days. TPA was measured in 4-fold diluted cell culture supernatants of unstimulated and PHA stimulated PBMC. Measured values were interpolated from the TPA Standard Curve diluted in Sample Diluent NS and DF corrected. Mean +/-SD, n=2, are graphed.

  • Serum from 10 apparently healthy male donors was measured in duplicate. The mean TPA concentration was determined to be 2,741 pg/mL with a range of 1,836- 4,012 pg/mL in male donors.

  • The concentrations of TPA were measured in duplicate and interpolated from the TPA standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TPA concentration was determined to be 4,710 pg/mL in serum and 4,270 pg/mL in plasma (heparin).

Protocols

References

ab190812 has not yet been referenced specifically in any publications.

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