Recombinant
RabMAb

Recombinant Anti-PAI1 antibody [EPR21850-82] (ab222754)

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Overview

  • Product name

    Anti-PAI1 antibody [EPR21850-82]
    See all PAI1 primary antibodies

  • Description

    Rabbit monoclonal [EPR21850-82] to PAI1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, ICC/IF, Flow Cyt, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human PAI1 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P05121

  • Positive control

    • WB: HepG2, HUVEC and Hepa1-6 whole cell lysate. Human, mouse and rat placenta lysate. Human liver lysate. Serum starved NIH/3T3 treated with TGF beta1 supernatant lysate. Serum starved NIH/3T3 treated with TGF beta1 and Brefeldin A whole cell lysate. ICC/IF: HUVEC cells. Serum starved NIH/3T3 treated with TGF beta and Brefeldin A cells. FC: Serum starved NIH/3T3 treated with TGF beta and Brefeldin A 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab222754 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
IP 1/30.
ICC/IF 1/100.
Flow Cyt 1/60.
WB 1/1000. Predicted molecular weight: 45 kDa.

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

    see all

  • Alternative names

    • Clade E antibody
    • Endothelial plasminogen activator inhibitor antibody
    • Nexin antibody
    • Nexin plasminogen activator inhibitor type 1 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
    • Serine proteinase inhibitor clade E member 1 antibody
    • serpin 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

  • Western blot - Anti-PAI1 antibody [EPR21850-82] (ab222754)
    Western blot - Anti-PAI1 antibody [EPR21850-82] (ab222754)
    All lanes : Anti-PAI1 antibody [EPR21850-82] (ab222754) at 1/1000 dilution

    Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 2 : Human liver lysate
    Lane 3 : Mouse placenta lysate
    Lane 4 : Rat placenta lysate
    Lane 5 : Hepa1-6 (Mouse hepatoma epithelial cell line) whole cell lysate
    Lane 6 : Human placenta lysate
    Lane 7 : HUVEC (Human umbilical vein endothelial cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lanes 1-5 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
    Lanes 6-7 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 45 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1-4 and 6: 3 minutes; Lane 5: 37 seconds: Lane 7: 8 seconds.

    PAI1 forms complex with its target protease, t-PA (lane 7). The molecular mass observed is consistent with what has been described in the literature (PMID 21596853).

    Lanes 6 and 7 were developed with a high sensitivity ECL substrate.

  • Immunocytochemistry/ Immunofluorescence - Anti-PAI1 antibody [EPR21850-82] (ab222754)
    Immunocytochemistry/ Immunofluorescence - Anti-PAI1 antibody [EPR21850-82] (ab222754)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HUVEC (Human umbilical vein endothelial cell line) cells labeling PAI1 with ab222754 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution (green). Confocal image showing cytoplasmic staining in HUVEC cell line. The nuclear counter stain is DAPI (blue).
    Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
    The negative control is the secondary antibody only.

  • Flow Cytometry - Anti-PAI1 antibody [EPR21850-82] (ab222754)
    Flow Cytometry - Anti-PAI1 antibody [EPR21850-82] (ab222754)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized NIH/3T3 cells serum-starved for 4 hours, treated with TGF-β (10 ng/ml) for 3 hours, and then with TGF-β (10 ng/ml) and BFA (300 ng/ml) together for 18 hours (Red) / Untreated control (Green) labeling PAI1 with ab222754 at 1/60 (red/green) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

    The expression of PAI-1 is induced by TGF-β in NIH/3T3 cell line (PMID 17890327).

  • Immunoprecipitation - Anti-PAI1 antibody [EPR21850-82] (ab222754)
    Immunoprecipitation - Anti-PAI1 antibody [EPR21850-82] (ab222754)

    PAI1 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell line) whole cell lysate with ab222754 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab222754 at 1/1,000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution.
    Lane 1: HepG2 whole cell lysate 10 µg (Input).
    Lane 2: ab222754 IP in HepG2 whole cell lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab222754 in HepG2 whole cell lysate (-).
    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 3 minutes.

  • Western blot - Anti-PAI1 antibody [EPR21850-82] (ab222754)
    Western blot - Anti-PAI1 antibody [EPR21850-82] (ab222754)
    All lanes : Anti-PAI1 antibody [EPR21850-82] (ab222754) at 1/1000 dilution

    Lane 1 : NIH/3T3 (Mouse embryonic fibroblast cell line) serum-starved for 4 hours, whole cell lysate
    Lane 2 : NIH/3T3 serum-starved for 4 hours then treated with 10 ng/ml TGF ß1 (ab50036) for 3 hours, then with 10 ng/ml TGF ß1 (ab50036) and 300 ng/ml Brefeldin A (BFA) together for 18 hours, whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 45 kDa


    Exposure time: 32 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression of PAI-1 is induced by TGF-β in the NIH/3T3 cell line (PMID 17890327). The 110 kDa band likely represents PAI-1 in complex with its target protease, t-PA (PMID 21596853).

    The blot was developed with a high sensitivity ECL substrate.

  • Western blot - Anti-PAI1 antibody [EPR21850-82] (ab222754)
    Western blot - Anti-PAI1 antibody [EPR21850-82] (ab222754)
    All lanes : Anti-PAI1 antibody [EPR21850-82] (ab222754) at 1/1000 dilution

    Lane 1 : NIH/3T3 (Mouse embryonic fibroblast cell line) serum-starved for 18 hours, then collected the supernatant lysate
    Lane 2 : NIH/3T3 serum-starved for 18 hours then treated with 10 ng/ml TGF ß1 (ab50036) for 24 hours, then collected the supernatant lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 45 kDa


    Exposure time: 26 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression of PAI-1 is induced by TGF-β in the NIH/3T3 cell line (PMID 17890327).

    The blot was developed with a high sensitivity ECL substrate.

  • Immunocytochemistry/ Immunofluorescence - Anti-PAI1 antibody [EPR21850-82] (ab222754)
    Immunocytochemistry/ Immunofluorescence - Anti-PAI1 antibody [EPR21850-82] (ab222754)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling PAI1 with ab222754 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution (green). Confocal image showing the signal is increased in 4 hour serum-starved NIH/3T3 cells treated with TGF-β (10 ng/ml) for 3 hours, then with TGF-β (10 ng/ml) and BFA (300 ng/ml) together for 18 hours. The expression of PAI-1 is induced by TGF-β in the NIH/3T3 cell line (PMID 17890327). The nuclear counter stain is DAPI (blue).
    Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
    The negative control is the secondary antibody only.

References

ab222754 has not yet been referenced specifically in any publications.

Publishing research using ab222754? Please let us know so that we can cite the reference in this datasheet.

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