Human Tissue type Plasminogen Activator ELISA Kit (ab119563)

Overview

  • Product nameHuman Tissue type Plasminogen Activator ELISA Kit
    See all Tissue type Plasminogen Activator kits
  • Detection methodColorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 8 3.6%
    Inter-assay
    Sample n Mean SD CV%
    Overall 8 6.5%
  • Tests
    1 x 96 test
  • Sample type
    Cell culture supernatant, Serum
  • Assay typeSandwich (quantitative)
  • Sensitivity
    6 pg/ml
  • Range
    15.6 pg/ml - 1000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 98 77% - 109%

  • Assay durationMultiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Abcam’s Tissue type Plasminogen Activator Human in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for accurate quantitative measurement of Human Tissue type Plasminogen Activator concentrations in Cell culture supernatant and serum.

    Tissue type Plasminogen Activator specific antibody has been precoated onto 96-well plates. Standards and test samples are added to the wells along with an HRP-conjugated Tissue type Plasminogen Activator detection antibody and the microplate is then incubated at room temperature and unbound proteins are then washed away with a wash buffer. TMB is then added and catalyzed by HRP to produce a blue color product that subsequently changes to yellow after addition of acidic stop solution. The density of yellow coloration is directly proportional to the Tissue type Plasminogen Activator amount of sample captured on the plate.

  • Tested applicationsSuitable for: Sandwich ELISAmore details
  • PlatformMicroplate

Properties

  • Storage instructionsStore at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Assay Buffer Concentrate 1 x 5ml
    20X Wash Buffer Concentrate 1 x 50ml
    Adhesive Films 2 units
    HRP-Conjugate anti-human Tissue type Plasminogen Activator polyclonal antibody 1 x 70µl
    Human Tissue type Plasminogen Activator Standard lyophilized 2 vials
    Microplate coated with polyclonal antibody to Human Tissue type Plasminogen Activator (12 x 8 wells) 1 unit
    Sample Diluent 1 x 12ml
    Stop Solution (1M Phosphoric acid) 1 x 15ml
    TMB Substrate Solution 1 x 15ml
  • Research areas
  • FunctionConverts 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 specificitySynthesized 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 diseaseNote=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 similaritiesBelongs 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.
  • DomainBoth 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 localizationSecreted > 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 ab119563 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.

Human Tissue type Plasminogen Activator ELISA Kit images

  • Representative Standard Curve using ab119563.

Protocols

References for Human Tissue type Plasminogen Activator ELISA Kit (ab119563)

ab119563 has not yet been referenced specifically in any publications.

Product Wall

testing rhesus crossreactivity

Excellent Excellent 5/5 (Ease of Use)
Abreviews
The Factor tPA ELISA, using the Abcam human ELISA kit ab119563, lt GR257353-2, was performed by an experienced laboratory technician at an accredited university. The goal of the experiment was to analyze the linearity of dilutions of rhesus macaque EDTA-prepared plasma samples and serum.

Overall, the kit, like other Abcam ELISA kits, was easy to follow, clear in terms of directions, and brief yet thorough. The laboratory technician had no problems understanding any of the protocol’s steps.

Our experiment consisted of using EDTA-prepared Rhesus Macaque plasma samples from N=3 M. mulatta. Samples were stored prior to thaw at -80 degrees Celsius. Plasma and Serum samples were diluted 1:5, 1:10, 1:20, and 1:40 in the given diluent as duplicates. The rest of the assay was performed per protocol’s instructions.

Absorbance measurements of the diluted samples were very comparable between duplicates. Samples diluted at 1:5, 1:10, and 1:20 gave similar extrapolated results and seemed to be in a linear range.

We performed a spike recovery as follows: 30uL of 2000ng/mL protein (from resuspension of the std) was spiked into 90ul of diluent, plasma, or serum. In Diluent, the recovery was 110% at 1:5, 1:10, and 1:20. In Plasma, recovery was 70% at the 1:20 dilution, but not detectable at 1:10 or 1:5. In the serum, recovery ranged between 25% and 40% in the tested dilutions.

Overall, we would recommend this kit for its ease of use and positive crossreactivity with rhesus samples.
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Submitted Jun 01 2016

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