Overview

  • Product name
    Anti-PAI1 antibody [EPR17795]
    See all PAI1 primary antibodies
  • Description
    Rabbit monoclonal [EPR17795] to PAI1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, WB, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human PAI1 aa 100-200. The exact sequence is proprietary.
    Database link: P05121

  • Positive control
    • WB: Human PAI1 full length protein; HepG2 whole cell lysate; Human fetal liver and fetal spleen lysates. ICC/IF: HepG2 and HT1080 cells. IP: HepG2 whole cell lysate
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab187262 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
Flow Cyt Use at an assay dependent concentration.
WB 1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).
ICC/IF 1/500.
IP 1/40.

Target

  • Function
    This inhibitor acts as 'bait' for tissue plasminogen activator, urokinase, and protein C. Its rapid interaction with TPA may function as a major control point in the regulation of fibrinolysis.
  • Tissue specificity
    Found in plasma and platelets and in endothelial, hepatoma and fibrosarcoma cells.
  • Involvement in disease
    Defects in SERPINE1 are the cause of plasminogen activator inhibitor-1 deficiency (PAI-1D) [MIM:613329]. It is a hematologic disorder characterized by increased bleeding after trauma, injury, or surgery. Affected females have menorrhagia. The bleeding defect is due to increased fibrinolysis of fibrin blood clots due to deficiency of plasminogen activator inhibitor-1, which inhibits tissue and urinary activators of plasminogen.
    Note=High concentrations of SERPINE1 seem to contribute to the development of venous but not arterial occlusions.
  • Sequence similarities
    Belongs to the serpin family.
  • Post-translational
    modifications
    Inactivated by proteolytic attack of the urokinase-type (u-PA) and the tissue-type (TPA), cleaving the 369-Arg-
    -Met-370 bond.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • Clade E antibody
    • Endothelial plasminogen activator inhibitor antibody
    • Nexin antibody
    • PAI 1 antibody
    • PAI antibody
    • PAI-1 antibody
    • PAI1_HUMAN antibody
    • PLANH1 antibody
    • Plasminogen activator inhibitor 1 antibody
    • Plasminogen activator inhibitor type 1 antibody
    • Serine (or cysteine) proteinase inhibitor antibody
    • Serine (or cysteine) proteinase inhibitor clade E (nexin plasminogen activator inhibitor type 1) member 1 antibody
    • Serpin E1 antibody
    • Serpin peptidase inhibitor clade E (nexin plasminogen activator inhibitor type 1) member 1 antibody
    • Serpin peptidase inhibitor clade E antibody
    • Serpine 1 antibody
    • SERPINE1 antibody
    see all

Images

  • All lanes : Anti-PAI1 antibody [EPR17795] (ab187262) at 1/1000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling PAI1 with ab187262 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm staining on HepG2 cells was observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab187262 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

  • ab187262 staining PAI1 in the human cell line HepG2 (human hepatocellular carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/60. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • PAI1 was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell lysate with ab187262 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab187262 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.

    Lane 1: HepG2 whole cell lysate 10 µg (Input). Lane 2: ab187262 IP in HepG2 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab187262 in HepG2 whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT1080 (Human fibrosarcoma cells) cells labeling PAI1 with ab187262 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm staining on HT1080 cells was observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab187262 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

  • Anti-PAI1 antibody [EPR17795] (ab187262) at 1/5000 dilution + HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 45 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-PAI1 antibody [EPR17795] (ab187262) at 1/5000 dilution + Human PAI1 full length protein at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 45 kDa
    Observed band size: 70 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

References

This product has been referenced in:
  • Xu J  et al. Epithelial-mesenchymal transition induced PAI-1 is associated with prognosis of triple-negative breast cancer patients. Gene 670:7-14 (2018). WB ; Human . Read more (PubMed: 29802992) »
  • Fang H  et al. PAI-1 induces Src inhibitor resistance via CCL5 in HER2-positive breast cancer cells. Cancer Sci 109:1949-1957 (2018). Read more (PubMed: 29601121) »
See all 2 Publications for this product

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