Anti-PAI1 antibody (ab66705)
Key features and details
- Rabbit polyclonal to PAI1
- Suitable for: IHC-P, ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
-
Product name
Anti-PAI1 antibody
See all PAI1 primary antibodies -
Description
Rabbit polyclonal to PAI1 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Horse, Cow, Pig -
Immunogen
Synthetic peptide corresponding to Human PAI1 aa 100-200 conjugated to keyhole limpet haemocyanin.
(Peptide available asab66704) -
Positive control
- This antibody gave a positive signal in the following Lysates: WB: HUVEC whole cell lysate and Human Aortic Endothelial Cell Lysate. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal kidney. ICC/IF: HeLa cell line.
-
General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
-
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
-
Related Products
- Dynasore, dynamin inhibitor (ab120192)
- Dynole® 34-2, dynamin I and dynamin II inhibitor (ab120463)
- Dynole® 31-2, Negative control for Dynolereg 34-2 (ab120464)
- MiTMAB™, dynamin I and dynamin II inhibitor (ab120466)
- OcTMAB™, dynamin I and dynamin II inhibitor (ab120467)
- BAPTA-AM, Ca2+ chelator (ab120503)
- Dyngo® 4a, Novel, highly potent dynamin inhibitor (ab120689)
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab66705 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 |
---|---|---|
IHC-P | (4) |
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
ICC/IF |
Use a concentration of 1 - 5 µg/ml.
|
|
WB | (3) |
Use a concentration of 1 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).
|
Notes |
---|
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
Use a concentration of 1 - 5 µg/ml. |
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 45 kDa (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
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
-
Database links
- Entrez Gene: 5054 Human
- Entrez Gene: 18787 Mouse
- Entrez Gene: 396945 Pig
- Entrez Gene: 24617 Rat
- Omim: 173360 Human
- SwissProt: P05121 Human
- SwissProt: P22777 Mouse
- SwissProt: P79335 Pig
see all -
Alternative names
- Clade E antibody
- Endothelial plasminogen activator inhibitor antibody
- Nexin antibody
see all
Images
-
All lanes : Anti-PAI1 antibody (ab66705) at 1 µg/ml
Lane 1 : HUVEC (Human Umbilical Vein Endothelial Cell) Whole Cell Lysate
Lane 2 : Human Aortic Endothelial Cell Lysate (HAEC)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 45 kDa
Observed band size: 45 kDa
Exposure time: 4 minutesThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab66705 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
-
All lanes : Anti-PAI1 antibody (ab66705) at 1 µg/ml
Lane 1 : HUVEC (Human Umbilical Vein Endothelial Cell) Whole Cell Lysate
Lane 2 : Human Aortic Endothelial Cell Lysate (HAEC)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 45 kDa
Observed band size: 42,45 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutesThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab66705 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
-
ab66705 staining PAI1 in MCF-7 cells treated with splitomicin (ab141120), by ICC/IF. Increase in PAI1 expression correlates with increased concentration of splitomicin, as described in literature.
The cells were incubated at 37°C for 48 hours in media containing different concentrations of ab141120 (splitomicin) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab66705 (1 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue -
IHC image of PAI1 staining in Human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab66705, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
-
ab66705 staining PAI1 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with the antibody ab66705 at 1µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
-
ab66705 staining PAI1 in HeLa cells treated with dynasore (ab120192), by ICC/IF. Increase in PAI1 expression correlates with increased concentration of dynasore, as described in literature.
The cells were incubated at 37°C for 6h in media containing different concentrations of ab120192 (dynasore) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab66705 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab66705 staining PAI1 in HeLa cells treated with Dyngo-4a™ (ab120689), by ICC/IF. Increase in PAI1 expression correlates with increased concentration of Dyngo-4a™, as described in literature.
The cells were incubated at 37°C for 6h in media containing different concentrations of ab120689 (Dyngo-4a™) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab66705 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab66705 staining PAI1 in HeLa cells treated with dynole-34-2™ (ab120463), by ICC/IF. Increase in PAI1 expression correlates with increased concentration of dynole-34-2™, as described in literature.
The cells were incubated at 37°C for 24h in media containing different concentrations of ab120463 (dynole-34-2™) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab66705 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab66705 staining PAI1 in HeLa cells treated with iminodyn-22™ (ab120461), by ICC/IF. Increase in PAI1 expression correlates with increased concentration of iminodyn-22™, as described in literature.
The cells were incubated at 37°C for 48h in media containing different concentrations of ab120461 (iminodyn-22™) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab66705 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab66705 staining PAI1 in HeLa cells treated with MiTMAB™ (ab120466), by ICC/IF. Increase in PAI1 expression correlates with increased concentration of MiTMAB™, as described in literature.
The cells were incubated at 37°C for 24h in media containing different concentrations of ab12046 (MiTMAB™) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab66705 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab66705 staining PAI1 in HeLa cells treated with OcTMAB™ (ab120467), by ICC/IF. Increase in PAI1 expression correlates with increased concentration of OcTMAB™, as described in literature.
The cells were incubated at 37°C for 24h in media containing different concentrations of ab120467 (OcTMAB™) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab66705 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab66705 staining PAI1 in HeLa cells treated with dynole-31-2™ (ab120464), by ICC/IF. No change in PAI1 expression with increased concentration of dynole-31-2™ (negative control for dynole 34-2™ (ab120463), as described in literature.
The cells were incubated at 37°C for 6h in media containing different concentrations of (ab120464 (dynole-31-2™) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab66705 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab66705 staining PAI1 in HepG2 cells treated with BAPTA sodium salt (ab120449), by ICC/IF. Increase in PAI1 expression correlates with increased concentration of BAPTA sodium salt, as described in literature.
The cells were incubated at 37°C for 4 hours in media containing different concentrations of ab120449 (BAPTA sodium salt) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab66705 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab66705 staining PAI1 in HepG2 cells treated with BAPTA-AM (ab120503), by ICC/IF. Increase in PAI1 expression correlates with increased concentration of BAPTA-AM, as described in literature.
The cells were incubated at 37°C for 4 hours in media containing different concentrations of ab120503 (BAPTA-AM) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab66705 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (71)
ab66705 has been referenced in 71 publications.
- Lottin M et al. Molecular Landscape of the Coagulome of Oral Squamous Cell Carcinoma. Cancers (Basel) 14:N/A (2022). PubMed: 35053621
- Irshad K et al. Upregulation of Atypical Cadherin FAT1 Promotes an Immunosuppressive Tumor Microenvironment via TGF-β. Front Immunol 13:813888 (2022). PubMed: 35720420
- Zeitlmayr S et al. TRPM7 restrains plasmin activity and promotes transforming growth factor-β1 signaling in primary human lung fibroblasts. Arch Toxicol 96:2767-2783 (2022). PubMed: 35864199
- Zhou Z et al. Connexin 43 overexpression induces lung cancer angiogenesis in vitro following phosphorylation at Ser279 in its C-terminus. Oncol Lett 24:293 (2022). PubMed: 35949588
- Kim WT et al. Secretory SERPINE1 Expression Is Increased by Antiplatelet Therapy, Inducing MMP1 Expression and Increasing Colon Cancer Metastasis. Int J Mol Sci 23:N/A (2022). PubMed: 36076991