Anti-TPA Tissue Plasminogen Activator antibody [2A153] (ab21049)

Overview

  • Product name
    Anti-TPA Tissue Plasminogen Activator antibody [2A153]
    See all TPA Tissue Plasminogen Activator primary antibodies
  • Description
    Mouse monoclonal [2A153] to TPA Tissue Plasminogen Activator
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Human, Xenopus laevis
  • Immunogen

    Recombinant full length protein: human tPA

  • Positive control
    • Recombinant human TPA
  • General notes

    This product used to be produced in ascites, lots up to and including GR248929-9 were produced in Ascites. All later lots are produced in TCS. 

Properties

Applications

Our Abpromise guarantee covers the use of ab21049 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
WB
IHC-FoFr
  • Application notes
    IHC-FoFr: Use at an assay dependent dilution (PMID 19074264).
    WB: Use at a concentration of 1 µg/ml. Use under non reducing conditions. Predicted molecular weight: 62 kDa.


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • 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
    • Database links
    • Alternative names
      • Alteplase antibody
      • DKFZp686I03148 antibody
      • Plasminogen activator tissue antibody
      • Plasminogen activator tissue type antibody
      • PLAT antibody
      • Reteplase antibody
      • t PA antibody
      • T Plasminogen Activator antibody
      • t-PA antibody
      • T-plasminogen activator antibody
      • Tissue plasminogen activator (t PA) antibody
      • Tissue type plasminogen activator antibody
      • Tissue-type plasminogen activator chain B antibody
      • tPA antibody
      • TPA_HUMAN antibody
      • TPA1 antibody
      see all

    References

    This product has been referenced in:
    • Bestman JE & Cline HT The RNA binding protein CPEB regulates dendrite morphogenesis and neuronal circuit assembly in vivo. Proc Natl Acad Sci U S A 105:20494-9 (2008). IHC-FoFr ; Xenopus laevis . Read more (PubMed: 19074264) »
    See 1 Publication for this product

    Customer reviews and Q&As

    1-6 of 6 Abreviews or Q&A

    Question
    Answer

    The exact epitope for Catalog # N63520M has not been mapped at this time, but it recognizes the heavy chain of Tissue Plasminogen Activator (tPA).

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rabbit Tissue sections (artery, spleen, small intestine, ovary,)
    Specification
    artery, spleen, small intestine, ovary,
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

    Abcam user community

    Verified customer

    Submitted Jul 22 2011

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (A549 cell line)
    Loading amount
    10 µg
    Specification
    A549 cell line
    Gel Running Conditions
    Reduced Denaturing (10% Bis-Tris gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C

    Abcam user community

    Verified customer

    Submitted Aug 15 2008

    Answer

    Please find below an extract from our protocol on WB (which can be found here https://www.abcam.com/assets/pdf/protocols/WB-beginner.pdf) for an explanation of non reducing conditions: E) Denaturation and reduction of the protein Antibodies typically recognize a small portion of the protein of interest (termed epitope) and this domain may reside within the 3D conformation of the protein. To enable access of the antibody to this portion it is necessary to unfold the protein, i.e. denature it. To do so, use a loading buffer with the anionic denaturing detergent sodium dodecyl sulfate (SDS), and boil the mixture at 95-100°C for 5 minutes. When SDS is used with proteins, all of the proteins become negatively charged by their attachment to the SDS anions. SDS denatures proteins by “wrapping around” the polypeptide backbone - and SDS binds to proteins fairly specifically in a mass ratio of 1.4:1. In so doing, SDS confers a negative charge to the polypeptide in proportion to its length - i.e., the denatured polypeptides become “rods” of negative charge clouds with equal charge or charge densities per unit length. It is usually necessary to reduce disulphide bridges in proteins before they adopt the random-coil configuration necessary for separation by size by using ß-mercaptoethanol or dithiothreitol (DTT). In denaturing SDS-PAGE separations, therefore, migration is determined not by intrinsic electrical charge of the polypeptide, but by molecular weight. SDS grade is of utmost importance: a protein stained background along individual gel tracts with indistinct or slightly distinct protein bands are indicative of old or poor quality SDS. Exceptions: 1) Certain antibodies recognize the native form (i.e. non-denatured form) of the protein. It is imperative in those circumstances to run a Western Blot in non-denaturing conditions. 2) Certain antibodies only recognize protein in its non-reduced form i.e. in an oxidized form (particularly on cysteine residues) and the reducing agents ß-mercaptoethanol and DTT must be removed from the traditional loading buffer and migration buffer (non reducing conditions). In summary… Non reduced: Loading Buffer: With SDS, No ß-mercaptoethanol, No DTT. Migration buffer: With SDS. I have asked the source of the antibody for the recommended dilution in WB, as soon as I receive it I will forward it to you,

    Read More

    Answer

    Thank you for your enquiry. Monoclonal Antibody to Human Tissue Plasminogen Activator(tPA), ab21049, was tested with recombinant human TPA in Western blot. I hope this information helps. Please do not hesitate to contact us again if you need anything further.

    Read More

    Answer

    Thank you for your enquiry. We have not seen this pattern in Western blot. The two bands you are seeing could be two closely spaced glycosylated forms of the TPA. I hope this information helps. Please do not hesitate to contact us if you need anything further.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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