Recombinant Anti-PAI1 antibody [EPR17272-21] (ab182973)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17272-21] to PAI1
- Suitable for: Flow Cyt (Intra), WB, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-PAI1 antibody [EPR17272-21]
See all PAI1 primary antibodies -
Description
Rabbit monoclonal [EPR17272-21] to PAI1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A549, HUVEC, Hepa1-6 and HepG2 whole cell lysates; mouse lung (7 days supernatant), mouse placenta, rat placenta, rat lung. ICC/IF: Hepa1-6 cells. Flow Cyt (intra): Hepa1-6 cells.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17272-21 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab182973 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 |
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Flow Cyt (Intra) |
1/60.
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WB |
1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).
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ICC/IF |
1/100.
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Notes |
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Flow Cyt (Intra)
1/60. |
WB
1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa). |
ICC/IF
1/100. |
Target
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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
modificationsInactivated 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
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Database links
- Entrez Gene: 5054 Human
- Entrez Gene: 18787 Mouse
- Entrez Gene: 24617 Rat
- Omim: 173360 Human
- SwissProt: P05121 Human
- SwissProt: P22777 Mouse
- SwissProt: P20961 Rat
- Unigene: 414795 Human
see all -
Alternative names
- Clade E antibody
- Endothelial plasminogen activator inhibitor antibody
- Nexin antibody
see all
Images
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All lanes : Anti-PAI1 antibody [EPR17272-21] (ab182973) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : SERPINE1 knockout A549 cell lysate
Lane 3 : HUVEC cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 45 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?Lanes 1 - 3: Merged signal (red and green). Green - ab182973 observed at 48 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab182973 was shown to react with PAI1 in wild-type A549 cells in Western blot. The band observed in the edited lysate lane above 45 kDa is likely to represent SERPINE1 with an insertion. This has not been investigated further. Wild-type A549 and SERPINE1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab182973 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Hepa1-6 (mouse hepatoma epithelial cell line) cells labeling PAI1 with ab182973 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Hepa1-6 cells.
The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
-ve control: PBS only, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution. -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methonol-permeabilized Hepa1-6 (mouse hepatoma epithelial cell line) cells labeling PAI1 with ab182973 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-PAI1 antibody [EPR17272-21] (ab182973) at 1/1000 dilution
Lane 1 : Rat placenta tissue lysate
Lane 2 : Rat lung tissue lysate
Lane 3 : Rat liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 45 kDa
Observed band size: 45 kDa
Exposure time: 20 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST.
The expression level of mouse and rat PAI1 may be low in normal liver tissue (PMID: 21898503). This antibody detects no bands in rat liver.
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All lanes : Anti-PAI1 antibody [EPR17272-21] (ab182973) at 1/1000 dilution
Lane 1 : Mouse placenta tissue lysate
Lane 2 : Mouse liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 45 kDa
Observed band size: 45 kDaBlocking/Diluting buffer and concentration: 5% NFDM/TBST.
The expression level of mouse and rat PAI1 may be low in normal liver tissue (PMID: 21898503). This antibody detects a 37kDa extra band without BOI in mouse liver.
Exposure time:
Lane 1 : 3.25 seconds
Lane 2 : 180 seconds -
All lanes : Anti-PAI1 antibody [EPR17272-21] (ab182973) at 1/1000 dilution
Lane 1 : Hepa1-6 (mouse hepatoma epithelial cell line) whole cell lysate
Lane 2 : HepG2 (human liver hepatocellular carcinoma cell line) 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
Developed using the ECL technique.
Predicted band size: 45 kDa
Observed band size: 45 kDaBlocking and dilution buffer: 5% NFDM/TBST
Exposure times.
Lane 1: 1 second
Lane 2: 3 seconds
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Anti-PAI1 antibody [EPR17272-21] (ab182973) at 1/1000 dilution + Mouse lung (7 days supernatant) at 10 µl
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 45 kDa
Observed band size: 45 kDa
Exposure time: 3 secondsBlocking and dilution buffer: 5% NFDM/TBST
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All lanes : Anti-PAI1 antibody [EPR17272-21] (ab182973) at 1/5000 dilution
Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) grown in serum-free media for 18 hours, whole cell lysate
Lane 2 : HepG2 grown in serum-free media for 18 hours, then treated with 10ng/ml TGFß for 24 hours, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 45 kDa
Observed band size: 45 kDa
Exposure time: 180 secondsBlocking and diluting buffer: 5% NFDM/TBST
This image was performed on a BIO-RAD® ChemiDoc™ MP instrument. The expression profile observed is consistent with what has been described in the literature (PMID: 20519507).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (7)
ab182973 has been referenced in 7 publications.
- Bruno MEC et al. Visceral fat-specific regulation of plasminogen activator inhibitor-1 in aged septic mice. J Cell Physiol 237:706-719 (2022). PubMed: 34369600
- Morishita M et al. Characterization of mouse embryonic fibroblasts derived from Rassf6 knockout mice shows the implication of Rassf6 in the regulation of NF-κB signaling. Genes Cells 26:999-1013 (2021). PubMed: 34652874
- Lucas JH et al. Multi-Walled Carbon Nanotubes (MWCNTs) Cause Cellular Senescence in TGF-ß Stimulated Lung Epithelial Cells. Toxics 9:N/A (2021). PubMed: 34205339
- Zwischenberger BA et al. Adipose-Derived Inflammatory and Coagulant Mediators in Patients With Sepsis. Shock 55:596-606 (2021). PubMed: 32496420
- Zheng D et al. Pyruvate Kinase M2 Tetramerization Protects against Hepatic Stellate Cell Activation and Liver Fibrosis. Am J Pathol 190:2267-2281 (2020). PubMed: 32805235
- Wang Q et al. E-cigarette-induced pulmonary inflammation and dysregulated repair are mediated by nAChR a7 receptor: role of nAChR a7 in SARS-CoV-2 Covid-19 ACE2 receptor regulation. Respir Res 21:154 (2020). PubMed: 32552811
- Wang Q et al. E-cigarette-Induced Pulmonary Inflammation and Dysregulated Repair are Mediated by nAChR a7 Receptor: Role of nAChR a7 in ACE2 Covid-19 receptor regulation. Res Sq N/A:N/A (2020). PubMed: 32702718