Recombinant Anti-PAI1 antibody [EPR17795] (ab187262)
![Western blot - Anti-PAI1 antibody [EPR17795] (ab187262)](https://www.abcam.com/ps/products/187/ab187262/Images/ab187262-454783-anti-pai1-antibody-epr17795-western-blot-wildtype-a549-serpine1-knockout-a549-huvec-hek293.jpg "Western blot - Anti-PAI1 antibody [EPR17795] (ab187262)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17795] to PAI1
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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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 (Intra), WB, ICC/IF, IPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human PAI1 full length protein; A549, HUVEC, HepG2 whole cell lysate; Human fetal liver and fetal spleen lysates. ICC/IF: HepG2 and HT1080 cells. IP: HepG2 whole cell lysate
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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.
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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17795 -
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 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 (Intra) |
Use at an assay dependent concentration.
|
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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 |
1/500.
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| IP |
1/40.
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| Notes |
|---|
|
Flow Cyt (Intra)
Use at an assay dependent concentration. |
|
WB
1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa). |
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ICC/IF
1/500. |
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IP
1/40. |
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
- Omim: 173360 Human
- SwissProt: P05121 Human
- Unigene: 414795 Human
- Unigene: 713079 Human
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Alternative names
- Clade E antibody
- Endothelial plasminogen activator inhibitor antibody
- Nexin antibody
see all
Images
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Western blot - Anti-PAI1 antibody [EPR17795] (ab187262)All lanes : Anti-PAI1 antibody [EPR17795] (ab187262) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : SERPINE1 knockout A549 cell lysate
Lane 3 : HUVEC cell lysate
Lane 4 : HEK-293 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 - 4: Merged signal (red and green). Green - ab187262 observed at 48 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab187262 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 ab187262 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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![Immunocytochemistry/ Immunofluorescence - Anti-PAI1 antibody [EPR17795] (ab187262)](https://www.abcam.com/ps/products/187/ab187262/Images/ab187262-244482-anti-pai1-antibody-epr17795-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-PAI1 antibody [EPR17795] (ab187262)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. -
![Flow Cytometry (Intracellular) - Anti-PAI1 antibody [EPR17795] (ab187262)](https://www.abcam.com/ps/products/187/ab187262/Images/ab187262-268977-anti-pai1-antibody-epr17795-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-PAI1 antibody [EPR17795] (ab187262)ab187262 staining PAI1in the human cell line HepG2 (human hepatocellular carcinoma) by intracellular 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)
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![Immunoprecipitation - Anti-PAI1 antibody [EPR17795] (ab187262)](https://www.abcam.com/ps/products/187/ab187262/Images/ab187262-244480-anti-pai1-antibody-epr17795-immunoprecipitation.jpg)
Immunoprecipitation - Anti-PAI1 antibody [EPR17795] (ab187262)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.
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![Western blot - Anti-PAI1 antibody [EPR17795] (ab187262)](https://www.abcam.com/ps/products/187/ab187262/Images/ab187262-244483-anti-pai1-antibody-epr17795-western-blot.jpg)
Western blot - Anti-PAI1 antibody [EPR17795] (ab187262)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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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![Immunocytochemistry/ Immunofluorescence - Anti-PAI1 antibody [EPR17795] (ab187262)](https://www.abcam.com/ps/products/187/ab187262/Images/ab187262-244481-anti-pai1-antibody-epr17795-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-PAI1 antibody [EPR17795] (ab187262)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. -
![Western blot - Anti-PAI1 antibody [EPR17795] (ab187262)](https://www.abcam.com/ps/products/187/ab187262/Images/ab187262-244484-anti-pai1-antibody-epr17795-western-blot.jpg)
Western blot - Anti-PAI1 antibody [EPR17795] (ab187262)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 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-PAI1 antibody [EPR17795] (ab187262)](https://www.abcam.com/ps/products/187/ab187262/Images/ab187262-244485-anti-pai1-antibody-epr17795-western-blot.jpg)
Western blot - Anti-PAI1 antibody [EPR17795] (ab187262)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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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![Anti-PAI1 antibody [EPR17795] (ab187262)](https://www.abcam.com/ps/products/187/ab187262/Images/ab187262-8-benefits-of-recombinant-antibodies.png)
Anti-PAI1 antibody [EPR17795] (ab187262)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
ab187262 has been referenced in 2 publications.
- 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 . 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). PubMed: 29601121