Tissue type Plasminogen Activator Activity Assay Kit (Colorimetric, Human) (ab108905)

Overview

  • Product name

    Tissue type Plasminogen Activator Activity Assay Kit (Colorimetric, Human)
  • 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.

  • 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.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    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

  • 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.
  • 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

    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

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

Images

  • tPA measured in biological fluids showing activity (IU) per mL of tested sample

  • Time course of tPA measured in biological fluids showing optical densities

  • Standard curve (colorimetric) : mean of duplicates (+/-SD) with background readings subtracted

  • Change of absorbance per min. of various concentrations of tPA standard

  • O.D's of various dilutions of human plasma after 20hrs using ab108905

     

  • Tissue type Plasminogen Activator Chromogenic Activity Standard curve

Protocols

References

This product has been referenced in:

  • 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). Read more (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). Read more (PubMed: 26876003) »
See all 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Mouse plasma testing

Excellent Excellent 5/5 (Ease of Use)
Abreviews
This kit works fine for mouse plasma.

Abcam user community

Verified customer

Submitted Mar 18 2016

Answer

Unfortunately we do not sell components from this kit separately.

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