Technical datasheet
Description
Plasminogen activator inhibitor type-1 (PAI-1), a member of the serine protease inhibitor (serpin) superfamily, is an important protein in the regulation of fibrinolysis. PAI-1 is unique among the serpins because of its functional and conformational flexibility. PAI-1 is the most important physiological inhibitor of both tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA). Increased PAI-1 levels are associated with thrombotic events and is an established risk factor for cardiovascular diseases. The active conformation PAI-1 inhibits its target proteinases by the formation of a stable, inactive complex. Although PAI-1 is synthesized as an active molecule, it converts spontaneously to an inactive, latent form that can be partially reactivated by denaturing agents. In addition, a third conformation reacting as a non-inhibitory substrate towards various target proteinases has been identified.
The epitope of monoclonal antibody MA-33H1F7 is predominantly composed of three residues (Lys154/Glu130/Arg131), positioned virtually linearly in the three-dimensional structure. The epitope of the antibody does not cover the complete alpha-helix F and turn connecting alpha-helix F and beta-strand s3A, but is restricted to the hinge region between alpha-helix F and the main part of the PAI-1 molecule.
The monoclonal antibody MA-33H1F7 is a ‘switching’ antibody, capable of inducing a non-inhibitory substrate form of PAI-1. It was shown to inhibit PAI-1 in a dose dependent manner.
Cross Reactivity
| Cross reactant |
Reactivity |
| Rat |
Yes |
| Mouse |
Yes |
|
|
Immunogen
Human PAI-1/t-PA complex
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin.
Species
Mouse IgG1
Application
|
|
F
|
FC
|
FS1,2,4,6
|
IA1
|
IF
|
IP
|
P
|
W5
|
|
Yes
|
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|
●
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●
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●
|
|
No
|
|
|
|
|
|
|
|
|
|
N.D.
|
●
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●
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|
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●
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●
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●
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N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot
Application notes
W: A non-reduced sample treatment and SDS-Page was used. The band size is 52 kDa (Ref.5).
FS: Antibody MA-33H1F7 functions as an antagonist. The antibody was incubated with active PAI-1 and residual activity was measured by a functional assay (Ref.1).
Use
For Western blotting, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50. For functional studies, dilutions have to be optimized in user’s experimental setting.
Aliases
PAI-1, endothelial plasminogen activator inhibitor, serpin E1
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least one year. The exact expiry date is indicated on the label.
References
1. Debrock, S et al; Neutralization of plasminogen activator inhibitor-1 inhibitory properties: identification of two different mechanisms. Biochim Biophys Acta 1997, 1337: 257
2. Berry, C et al; Antithrombotic activity of a monoclonal antibody inducing the substrate form of plasminogen activator inhibitor type 1 in rat models of venous and arterial thrombosis. Br J Pharm 1998, 125: 29
3. Bijnens, A et al; Importance of the hinge region between alpha-helix F and the main part of serpins, based upon identification of the epitope of plasminogen activator inhibitor type 1 neutralizing antibodies. J Biol Chem 2000, 275: 6375
4. Rupin, A et al; Inactivation of plasminogen activator inhibitor-1 accelerates thrombolysis of a platelet-rich thrombus in rat mesenteric arterioles. Thromb Haemst 2001, 86: 1528
5. Sironi, L et al; Effect of valsartan on angiotensin ll-induced plasminogen activator inhibitor-1 biosynthesis in arterial smooth muscle cells. J Am Heart Ass 2001, 37: 961
6. Komissarov, A et al; Redirection of the reaction between activated protein C and a serpin to the substrate pathway. Thromb Res. 2008, 122: 397
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.
Also available
References
1. Debrock, S et al; Neutralization of plasminogen activator inhibitor-1 inhibitory properties: identification of two different mechanisms. Biochim Biophys Acta 1997, 1337: 257
2. Berry, C et al; Antithrombotic activity of a monoclonal antibody inducing the substrate form of plasminogen activator inhibitor type 1 in rat models of venous and arterial thrombosis. Br J Pharm 1998, 125: 29
3. Bijnens, A et al; Importance of the hinge region between alpha-helix F and the main part of serpins, based upon identification of the epitope of plasminogen activator inhibitor type 1 neutralizing antibodies. J Biol Chem 2000, 275: 6375
4. Rupin, A et al; Inactivation of plasminogen activator inhibitor-1 accelerates thrombolysis of a platelet-rich thrombus in rat mesenteric arterioles. Thromb Haemst 2001, 86: 1528
5. Sironi, L et al; Effect of valsartan on angiotensin ll-induced plasminogen activator inhibitor-1 biosynthesis in arterial smooth muscle cells. J Am Heart Ass 2001, 37: 961
6. Komissarov, A et al; Redirection of the reaction between activated protein C and a serpin to the substrate pathway. Thromb Res. 2008, 122: 397