uPA ELISA Kit (Human) (OKBB00267)

Catalog No: OKBB00267

Size:96 Tests

Human PLAU 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 uPA ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for uPA has been precoated onto 96-well plates. Standards(NS0, M1-L431) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for uPA 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 uPA amount of sample captured in plate.
• ELISA Kit Range: 62.5 pg/ml - 4,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	126	1895	2619	250	1568	2755
  St. Dev.	5.92	64.43	138.8	13.5	95.65	187.3
  %CV	4.69	3.4	5.29	5.4	6.10	6.79

• ELISA Kit Component:
  • Lyophilized recombinant human uPA/PLAU standard: 10 ng/tube x 2
  • Anti-human uPA/PLAU Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human PLAU 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: 62.5pg/ml-4000pg/ml
  Principle: Aviva’s human Human uPA/PLAU ELISA Kit was based on standard sandwich enzyme-linked immunesorbent
  assay technology. A monoclonal antibody from mouse specific for uPA/PLAU has been precoated onto 96-well plates. Standards (Expression system for standard: NSO; Immunogen sequence: M1-L431) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for uPA/PLAU 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 density of yellow is proportional to the Human uPA/PLAU 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,urokinase (PLAU, uPA) converts plasminogen to plasmin. Plasmin is involved in processing of amyloid precursor protein and degrades secreted and aggregated amyloid-beta, a hallmark of Alzheimer disease (AD).1 Urokinase has a molecular mass of about 54 kD and is composed of 2 disulfide-linked chains, A and B, of molecular masses 18 kD and 33 kD, respectively. It localized on 10q24. uPA facilitates cell migration by localizing proteolisys on the cell surface and by inducing intracellular signalling pathways. In human vascular smooth muscle cell (VSMC), uPA stimulates migration via the uPA receptor (uPAR) signalling complex containing TYK2 and phosphatidylinositol 3-kinase (PI3-K)
• 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: cell culture supernates, serum and plasma(heparin, EDTA)
• Sensitivity: <5 pg/ml
• Reference: 1. Finckh, U.; van Hadeln, K.; Muller-Thomsen, T.; Alberici, A.; Binetti, G.; Hock, C.; Nitsch, R. M.; Stoppe, G.; Reiss, J.;
  Gal, A. : Association of late-onset Alzheimer disease with a genotype of PLAU, the gene encoding urokinase-type
  plasminogen activator on chromosome 10q22.2. Neurogenetics 4: 213-217, 2003.
  2. Kiian, I.; Tkachuk, N.; Haller, H.; Dumler, I. : Urokinase-induced migration of human vascular smooth muscle cells
  requires coupling of the small GTPase RhoA and Rac1 to the Tyk2/PI3-K signalling pathway. Thromb. Haemost. 89:
  904-914, 2003.
• Specificity: Natural and recombinant human Upa
• Immunogen: Expression system for standard: NS0; Immunogen sequence: M1-L431
• Description: For quantitative detection of human uPA in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Target Info

• Gene Symbol: PLAU
• Gene Full Name: plasminogen activator, urokinase
• Alias Symbols: ATF, BDPLT5, Plasminogen activator urinary, PLAU, QPD, u PA, U-plasminogen activator, uPA, URK, UROK_HUMAN, Urokinase-type plasminogen activator chain B
• NCBI Gene Id: 5328
• Protein Name: Urokinase-type plasminogen activator
• Description of Target: Plasminogen activator,urokinase (PLAU, uPA) converts plasminogen to plasmin. Plasmin is involved in processing of amyloid precursor protein and degrades secreted and aggregated amyloid-beta, a hallmark of Alzheimer disease (AD). Urokinase has a molecular mass of about 54 kD and is composed of 2 disulfide-linked chains, A and B, of molecular masses 18 kD and 33 kD, respectively. It localized on 10q24. uPA facilitates cell migration by localizing proteolisys on the cell surface and by inducing intracellular signalling pathways. In human vascular smooth muscle cell (VSMC), uPA stimulates migration via the uPA receptor (uPAR) signalling complex containing TYK2 and phosphatidylinositol 3-kinase (PI3-K).
• Uniprot ID: P00749
• Protein Accession #: NP_001138503.1
• Nucleotide Accession #: NM_001145031.2