PAI-1 ELISA Kit (Human) (OKBB00347)

Catalog No: OKBB00347

Size:96 Tests

Human SERPINE1 ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human PAI-1 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for PAI-1 has been precoated onto 96-well plates. Standards(sf21, S22-P402) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for PAI-1 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human PAI-1 amount of sample captured in plate.
• ELISA Kit Range: 156 pg/ml - 10,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	1539	3736	6234	1637	3914	7335
  St. Dev.	76.95	235.4	405.21	104.8	293.55	601.47
  %CV	5	6.30	6.5	6.40	7.5	8.2

• ELISA Kit Component:
  • Lyophilized recombinant mouse C-MET standard: 10 ng/tube x 2
  • Anti-mouse C-MET Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-mouse C-MET antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 156pg/ml-10, 000pg/ml
  Principle: Aviva’s human PAI-1 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human PAI-1 specific-specific polyclonal antibodies were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human PAI-1 amount of sample captured in plate.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before using.
  Background: Plasminogen activator inhibitor-1 is the principal inhibitor of tissue plasminogen activator (tPA) and urokinase (uPA), the activators of plasminogen and hence fibrinolysis (the physiological breakdown of blood clots). It is a serine protease inhibitor (serpin) protein (SERPINE1). PAI-1 is mainly produced by the endothelium (cells lining blood vessels), but is also secreted by other tissue types, such as adipose tissue. The PAI-1 gene is located on the seventh chromosome (7q21.3-q22).1 Congenital deficiency of PAI-1 leads to a hemorrhagic diathesis (a tendency to hemorrhage). PAI-1 is present in increased levels in various disease states (such as a number of forms of cancer), as well as in obesity and the metabolic syndrome. It has been linked to the increased occurrence of thrombosis in patients with these conditions. Additionally, it has been shown to interact with ORM1.2 The standard product used in this kit is recombinant human PAI-1 with the molecular mass of 43KDa.
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: plasma, tissue lysates and cell culture supernates
• Sensitivity: <10 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Klinger, K. W.; Winqvist, R.; Andreasen, P. A.; Stuart, N.; Stanislovitis, P.; Schwartz, R.; Doherty, R.; Alitalo, K.;
  Blasi, F.; Dano, K. : Regional mapping of the plasminogen activator inhibitor type 1 gene to 7q21.3-q22 and
  assignment to the CF linkage group. (Abstract) Cytogenet. Cell Genet. 46: 640 only, 1987.
  2. Boncela, J; Papiewska I, Fijalkowska I, Walkowiak B, Cierniewski C S (Sep. 2001). "Acute phase protein alpha
  1-acid glycoprotein interacts with plasminogen activator inhibitor type 1 and stabilizes its inhibitory activity". J. Biol.
  Chem. (United States) 276 (38): 35305-11.
• Specificity: Natural and recombinant human PAI-1
• Immunogen: Expression system for standard: sf21; Immunogen sequence: S22-P402
• Description: For quantitative detection of human PAI-1 in cell culture supernatants, cell lysates, serum and plasma(heparin, EDTA, citrate ).

Target Info

• Gene Symbol: SERPINE1
• Gene Full Name: serpin peptidase inhibitor, clade E (nexin, plasminogen activator inhibitor type 1), member 1
• Alias Symbols: Endothelial plasminogen activator inhibitor, PAI, PAI-1, PAI1, PAI1_HUMAN, PLANH1, Plasminogen activator inhibitor 1, Serine (or cysteine) proteinase inhibitor clade E (nexin plasminogen activator inhibitor type 1) member 1, Serpin E1, Serpin peptidase inhibitor clade E (nexin plasminogen activator inhibitor type 1) member 1, SERPINE1
• NCBI Gene Id: 5054
• Protein Name: Plasminogen activator inhibitor 1
• Description of Target: Plasminogen activator inhibitor-1 is the principal inhibitor of tissue plasminogen activator (tPA) and urokinase (uPA), the activators of plasminogen and hence fibrinolysis (the physiological breakdown of blood clots). It is a serine protease inhibitor (serpin) protein (SERPINE1). PAI-1 is mainly produced by the endothelium (cells lining blood vessels), but is also secreted by other tissue types, such as adipose tissue. The PAI-1 gene is located on the seventh chromosome (7q21.3-q22).1 Congenital deficiency of PAI-1 leads to a hemorrhagic diathesis (a tendency to hemorrhage). PAI-1 is present in increased levels in various disease states (such as a number of forms of cancer), as well as in obesity and the metabolic syndrome. It has been linked to the increased occurrence of thrombosis in patients with these conditions. Additionally, it has been shown to interact with ORM1.2 The standard product used in this kit is recombinant human PAI-1 with the molecular mass of 43KDa.
• Uniprot ID: P05121
• Protein Accession #: NP_000593.1
• Nucleotide Accession #: NM_000602.4