Anti-Tissue Plasminogen Activator antibody [H27B6] (ab28374)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPA Tissue Plasminogen Activator antibody [H27B6] (ab28374)

    Key features and details

    • Mouse monoclonal [H27B6] to Tissue Plasminogen Activator
    • Suitable for: WB, ELISA, IHC-P
    • Reacts with: Mouse, Rat
    • Isotype: IgG

    Get better batch-to-batch reproducibility with a recombinant antibody

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    Anti-Tissue Plasminogen Activator antibody [EPR7232(2)] (ab157469)
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    • Success from the first experiment – confirmed specificity through extensive validation
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    Overview

    • Product name

      Anti-Tissue Plasminogen Activator antibody [H27B6]
      See all Tissue Plasminogen Activator primary antibodies

    • Description

      Mouse monoclonal [H27B6] to Tissue Plasminogen Activator

    • Host species

      Mouse

    • Tested applications

      Suitable for: WB, ELISA, IHC-Pmore details

    • Species reactivity

      Reacts with: Mouse, Rat

    • Immunogen

      Recombinant full length protein (Mouse)

    • Epitope

      protease domain (see Declerck et al)

    • Positive control

      • This antibody gave a positive result in IHC in the following FFPE tissue: Rat Normal Kidney.

    • General notes

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Properties

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab28374 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
    Use at an assay dependent concentration.
    ELISA
    Use at an assay dependent concentration.
    IHC-P
    Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
    Notes
    WB
    Use at an assay dependent concentration.
    ELISA
    Use at an assay dependent concentration.
    IHC-P
    Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    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

      Images

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPA Tissue Plasminogen Activator antibody [H27B6] (ab28374)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPA Tissue Plasminogen Activator antibody [H27B6] (ab28374)

        IHC image of TPA Tissue Plasminogen Activator staining in Rat Normal Kidney formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab28374, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

         

        For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      References (4)

      Publishing research using ab28374? Please let us know so that we can cite the reference in this datasheet.

      ab28374 has been referenced in 4 publications.

      • Cho MK  et al. Zinc-triggered induction of tissue plasminogen activator and plasminogen in endothelial cells and pericytes. Exp Neurobiol 22:315-21 (2013). WB . PubMed: 24465147
      • Hwang IY  et al. Zinc-triggered induction of tissue plasminogen activator by brain-derived neurotrophic factor and metalloproteinases. J Neurochem 118:855-63 (2011). WB . PubMed: 21615740
      • Cacquevel M  et al. Ageing and amyloid-beta peptide deposition contribute to an impaired brain tissue plasminogen activator activity by different mechanisms. Neurobiol Dis 27:164-73 (2007). PubMed: 17566751
      • Declerck PJ  et al. Generation of monoclonal antibodies against autologous proteins in gene-inactivated mice. J Biol Chem 270:8397-400 (1995). PubMed: 7536737

      Customer reviews and Q&As

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      1-3 of 3 Abreviews or Q&A

      Question

      Inquiry: We got multiple bands using this antibody and could not figure out which band is the one we want. The cell type we used is astroctyes from primary culture. 1. 35ug amount of protein was loaded. 2. Running 4-12%Gel (Invitrogen) 1and half hour at 130v and Transfer 3hrs at 80v. 3. Blocking 1 hour in 5%milk 4. 1:1000 Dilution of primary antibody, incubated overnight at 4C 5. 1:1000 Dilution of secondary antibody, incubated 1hr at RT 6. Using ECL to detect. 7. Washing 10mins×3 times after Primary and secondary antibody.

      Read More
      Answer

      I have just noticed that you were kind enough to include a western blot image on your initial email. I'm sorry to have missed that in my earlier email.

      After looking at your blot I think it might be helpful to reduce the antibody concentration of both the primary and secondary antibody. This may reduce many of these background bands. I recommend trying a 1:2000 dilution of the primary and a 1:10 000 dilution of the secondary to begin.

      I look forward to hearing if this advice proves useful.

      Read More

      Question

      Inquiry: We got multiple bands using this antibody and could not figure out which band is the one we want.

      The cell type we used is astroctyes from primary culture.
      1. 35ug amount of protein was loaded.
      2. Running 4-12%Gel (Invitrogen) 1and half hour at 130v and Transfer 3hrs at 80v.
      3. Blocking 1 hour in 5%milk
      4. 1:1000 Dilution of primary antibody, incubated overnight at 4C
      5. 1:1000 Dilution of secondary antibody, incubated 1hr at RT
      6. Using ECL to detect.
      7. Washing 10mins×3 times after Primary and secondary antibody.

      Read More
      Answer

      Thank you for your enquiry and for providing the protocol details.

      I think it would be helpful if you could provide an image of your western blot. When you have a chance, please send it along as an attachement.

      Thanks!

      Read More

      Question

      Can you please tell me if any of your Tissue Plasminogen Activator antibodies recognize only folded, active tPA or also unfolded, inactive tPA?

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      Answer

      The source of the antibody informed me " tPA does not have an active and a latent form. tPA is active in both single chain form and also once cleaved or the two chain form. If there is a latent or inactive form it is a new one on me. Your customer may be talking about a proform of tPA but, I am not familiar with this either. Our antibody for tPA has not been tested to this depth for reactivity regardless and therefore I cannot say if there is a any reactivity with the AB to folded or unfolded tPA." I hope this information will help,

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