Anti-TPA Tissue Plasminogen Activator antibody (ab14198)

Overview

  • Product name
    Anti-TPA Tissue Plasminogen Activator antibody
    See all TPA Tissue Plasminogen Activator primary antibodies
  • Description
    Goat polyclonal to TPA Tissue Plasminogen Activator
  • Host species
    Goat
  • Specificity
    There were no cross reactivities obtained with human uPA and PAI2.
  • Tested applications
    Suitable for: ELISA, IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Full length native protein (purified) (Human).

Properties

Applications

Our Abpromise guarantee covers the use of ab14198 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
ELISA Use at an assay dependent dilution.
IHC-Fr 1/50 - 1/100.
IHC-P 1/50. Pretreatment with 0.05% Pronase E for 20 minutes is recommended.

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

  • a14198 staining TPA Tissue Plasminogen Activator (A - blue) in Rat hippocampal/thalamic region tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde. Tissue was incubated in pre-block solution (10% normal hourse serum in antibody diluent (PBS + 0.1% Trition X-100)) for 1 hour. Samples were incubated with primary antibody (1/50 in PBS + pre-block solution) for 16 hours. B - trypsin (red), C - PAR2 (green), D - merge.

    TPA Tissue Plasminogen Actvator and Trypsin co-localise.

  • ab14198 at 1/50 staining rat brain hippocampus pyramidal neuron tissue sections by IHC-Fr. The tissue was blocked with horse serum and incubated for 16 hours with the antibody. An AMCA conjugated donkey antibody was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Mauprivez C  et al. Periosteum Metabolism and Nerve Fiber Positioning Depend on Interactions between Osteoblasts and Peripheral Innervation in Rat Mandible. PLoS One 10:e0140848 (2015). Read more (PubMed: 26509533) »
  • Granieri L  et al. A competition-based assay for the screening of species-specific antibiotics. J Antimicrob Chemother 64:62-8 (2009). IF ; Human . Read more (PubMed: 19401303) »
See all 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Brain, Hippocampus, pyramidal neurons)
Specification
Brain, Hippocampus, pyramidal neurons
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%

Mr. Rink-Jan Lohman

Verified customer

Submitted May 08 2007

Question

Could we get a detailed protocol from you, please. We have some general questions, the answers to which will enable us to investigate this matter as quickly as possible. ab14198 The tPA antibody was diluted by 1:(50~150), aliquoted and stored in 4? for use. 2. Please describe the problem (high background, no staining etc). Non-specific staining with high background stain:The paraffin enbedded uterine leiomyoma and normal smooth muscle was stain by this Ab, the result showed the high background and all the cells including smooth muscle, endometrial gland, endometrial stroma and vessels are all stained. 3. On what material are you testing the antibody in IHC? Species: uterus Cell Iine or tissue: paraffin-enbedded uterine leiomyoma 4. How did you fix the samples? Paraformaldehyde (10%) 5. For formalin-fixed paraffin embedded tissue: did you apply antigen retrieval step? Heat mediated technique 6. Blocking steps: For HRP detection method: did you block endogenous peroxidases: with H2O2 (3%, 20 min) How did you block the unspecific binding sites: goat serum For how long: 20 min 7. Primary antibody Specification (in which species was it raised against): human At what dilution(s) have you tested this antibody: 1:50~1:150 What dilution buffer was used: antibody dilution buffer Incubation time: overnight Incubation temperature: 4 What washing steps were done: wash by PBS for 3 times 8. Secondary antibody Specification (in which species was it raised against): Goat What dilution buffer was used: antibody dilution buffer Incubation time: 20 min Incubation temperature: 37 What washing steps were done: wash by PBS for 3 times Do you know whether the problems you are experiencing come from the secondary? No. 9. What detection method are you using? DAB0. Background staining Please provide an image of your staining 11. Which detection system did you use? Olympas BX41 12. Did you apply positive and negative controls along with the samples? Please specify.No. 13. Optimization attempts? How many times have you tried the IHC? 10 times Do you obtain the same results every time? Yes. What steps have you altered? use the goat serum to block.

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Answer

Thank you for your enquiry. I have been in contact with the originator of this antibody and have received the protocol that they used. Immunostraining Formalin-Fixed, Paraffin Embedded Tissues 1. Dewaxing: Xylene 5 min.- Xylene 5 min.- Xylene 5 min. 2. Rehydration: 100 % ethanol for 3 min.- 95 % ethanol for 3 min.- 70 % ethanol for 3 min. 3. Wash sections under running tap water for 5 minutes. 4. Wash sections in PBS. 5. Block endogenous peroxidase : 0.3 % Hydrogen Peroxid in absolute methanol (2ml of H202 30% in 198 ml methanol) for 30 minutes. (Cover the jar with foil, use stirring during incubation.) 6. Wash sections under running tap water for 10 minutes. 7. Tissue area with pap pen. 8. Cover the tissue area with 300µl of blocking solution, for 30 minutes at room temperature (blocking solution : 6ml PBS, 0.3 gm BSA, 120µl normal goat serum). 9. Shake off excess fluid (do not wash !). 10. Incubate with primary antibody at 4 °C overnight in moist chamber. 11. Rinse with PBS-T, then wash 3 times in PBS-T each for 5 minutes at room temp. 12. Incubate with biotinylated secondary antibody. Incubation for 30 minutes at room temp. 13. Rinse with PBS-T, then wash 3 times in PBS-T 3 x 5 minutes at room temp. 14. Incubate the sections with Vectastain ABC reagent for 30 minutes at room temp. (ABC reagent: 2 drops of A and 2 drops of B, in 5ml PBS, allow to stand 30 minutes at room temp. before use.) 15. Wash sections in PBS 3 x 5 minutes at room temp. 16. Develop with DAB for 5-10 minutes. 17. Wash under running tap water. 18. Counter stain with Mayer´s HX for 45 seconds, wash in warm tap water. 19. Dehydrate: 70% ethanol-95% ethanol-100% ethanol, 1minute each. 20. Wash in Xylene, mount, dry. Concentration of primary antibodies: Mouse monoclonal anti human******: 1: 2000 in PBS-T Concentration of secondary antibodies: Biotinylated goat anti-mouse antibody, SIGMA product No. B74001. Concentration was 1 : 400 in PBS-T. If problems for staining occur try treatement with incubating citrate buffer or microwavetreatement I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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