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Catalog No: OKBB00267
Size:96 Tests
Price: $583.00
SKU
OKBB00267
Availability: Domestic: within 1-2 week delivery | International: within 1-2 week delivery
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Datasheets/ManualsClick here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Product Info
Predicted Species ReactivityHuman
ApplicationELISA
ELISA Kit Detection MethodColorimetric, OD450 nm
ELISA Kit Duration~ 3 Hours
ELISA Kit PrincipleAviva'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 Range62.5 pg/ml - 4,000 pg/ml
ELISA Kit Reproducibility
Intra-Assay PrecisionInter-Assay Precision
Sample123123
Mean (pg/mL)1261895261925015682755
St. Dev.5.9264.43138.813.595.65187.3
%CV4.693.45.295.46.106.79
ELISA Kit Component
ComponentAmount
Lyophilized recombinant human uPA/PLAU standard10 ng/tube x 2
Anti-human uPA/PLAU Antibody Well Plate96 Wells
Sample diluent buffer30 mL
Biotinylated anti-human PLAU antibody130 uL, dilution 1:100
Antibody diluent buffer12 mL
Avidin-Biotin-Peroxidase Complex (ABC)130 uL, dilution 1:100
ABC diluent buffer12 mL
TMB color developing agent10 mL
TMB stop solution10 mL
Additional InformationRange: 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 StorageStore at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
Sample Typecell culture supernates, serum and plasma(heparin, EDTA)
Sensitivity<5 pg/ml
Reference1. 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.
SpecificityNatural and recombinant human Upa
ImmunogenExpression system for standard: NS0; Immunogen sequence: M1-L431
Gene SymbolPLAU
Gene Full Nameplasminogen activator, urokinase
Alias SymbolsATF, BDPLT5, Plasminogen activator urinary, PLAU, QPD, u PA, U-plasminogen activator, uPA, URK, UROK_HUMAN, Urokinase-type plasminogen activator chain B
NCBI Gene Id5328
Protein NameUrokinase-type plasminogen activator
Description of TargetPlasminogen 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 IDP00749
Protein Accession #NP_001138503.1
Nucleotide Accession #NM_001145031.2
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