Recombinant Anti-PAI1 antibody [EPR17796] - BSA and Azide free (ab250925)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17796] to PAI1 - BSA and Azide free
- Suitable for: ICC/IF, IP, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-PAI1 antibody [EPR17796] - BSA and Azide free
See all PAI1 primary antibodies -
Description
Rabbit monoclonal [EPR17796] to PAI1 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IP, WBmore details -
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
ab250925 is the carrier-free version of ab187263.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Clonality
Monoclonal -
Clone number
EPR17796 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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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 ab250925 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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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).
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Notes |
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ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa). |
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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All lanes : Anti-PAI1 antibody [EPR17796] (ab187263) 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?This data was developed using the same antibody clone in a different buffer formulation (ab187263).
Lanes 1 - 4: Merged signal (red and green). Green - ab187263 observed at 48 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab187263 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 ab187263 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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This data was developed using ab187263, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT1080 (Human fibrosarcoma cells) cells labeling PAI1 with ab187263 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: ab187263 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 [EPR17796] (ab187263) at 1/10000 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: 3 minutesThis data was developed using ab187263, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
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This data was developed using ab187263, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling PAI1 with ab187263 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: ab187263 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. -
All lanes : Anti-PAI1 antibody [EPR17796] (ab187263) 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 minutesThis data was developed using ab187263, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
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This data was developed using ab187263, the same antibody clone in a different buffer formulation.PAI1 was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell lysate with ab187263 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab187263 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: ab187263 IP in HepG2 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab187263 in HepG2 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 8 seconds. -
Anti-PAI1 antibody [EPR17796] (ab187263) 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 secondsThis data was developed using ab187263, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
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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Datasheet download
Certificate of Compliance
References (0)
ab250925 has not yet been referenced specifically in any publications.