ELISA Kit for Plasminogen Activator, Urokinase (uPA) Rattus norvegicus (Rat) Sandwich ELISA

PLAU; ATF; URK; UK; UP-A; Abbokinase; Urokinase-Type Plasminogen Activator

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  • ELISA Kit for Plasminogen Activator, Urokinase (uPA) Packages (Simulation)
  • ELISA Kit for Plasminogen Activator, Urokinase (uPA) Packages (Simulation)
  • ELISA Kit for Plasminogen Activator, Urokinase (uPA) Results demonstration
  • SEA140Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Plasminogen Activator, Urokinase (uPA) and the recovery rates were calculated by comparing the measured value to the expected amount of Plasminogen Activator, Urokinase (uPA) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-105 95
EDTA plasma(n=5) 78-99 90
heparin plasma(n=5) 82-98 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Plasminogen Activator, Urokinase (uPA) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Plasminogen Activator, Urokinase (uPA) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Plasminogen Activator, Urokinase (uPA) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 86-99% 80-92% 94-103% 78-91%
EDTA plasma(n=5) 87-102% 81-94% 85-92% 95-105%
heparin plasma(n=5) 79-98% 84-101% 90-98% 80-103%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Plasminogen Activator, Urokinase (uPA)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Plasminogen Activator, Urokinase (uPA). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Plasminogen Activator, Urokinase (uPA). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Plasminogen Activator, Urokinase (uPA), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Plasminogen Activator, Urokinase (uPA) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Oral administration of benzyl-isothiocyanate inhibits solid tumor growth and lung metastasis of 4T1 murine mammary carcinoma cells in BALB/c miceSpringerLink: f3w831267267p277
  • Decreased Hepatocyte Growth Factor (HGF) and Gamma Aminobutyric Acid (GABA) in Individuals with Obsessive-Compulsive Disorder (OCD)PubMed: PMC3762604
  • Downregulation of MACC1 inhibits invasion, migration and proliferation, attenuates cisplatin resistance and induces apoptosis in tongue squamous cell carcinomaPubmed:25421538
  • 1, 25-Dihydroxyvitamin D3 alleviates salivary adenoid cystic carcinoma progression by suppressing GPX1 expression through the NF-κB pathwayPubmed:26782341
  • Methylation of neurofilament light polypeptide promoter is associated with cell invasion and metastasis in NSCLCPubmed:26801564
  • Cytokine and Estrogen Stimulation of Endothelial Cells Augments Activation of the Surface-Bound Prekallikrein-High Molecular Weight Kininogen Complex: Implications for Hereditary Angioedema (HAE): 814article:S0091-6749(15)02643-3
  • A Study Protocol: Spinal Morphology, Physical Performance, Quality of Life and Biochemical Markers in Adults at Risk of Osteoporotic Fractures 59f7c85aaca272607e2d8d7a
  • Urokinase plasminogen activator secreted by cancer-associated fibroblasts induces tumor progression via PI3K/AKT and ERK signaling in esophageal squamous cell carcinomapubmed:28404945
  • Cytokine and estrogen stimulation of endothelial cells augments activation of the prekallikrein-high molecular weight kininogen complex: Implications for hereditary angioedema.pubmed:27826093
  • Effect of a synthetic inhibitor of urokinase plasminogen activator on the migration and invasion of human cervical cancer cells in vitroPubmed:29328476
  • Multi-omics analysis reveals the mechanisms of action and therapeutic regimens of traditional Chinese medicine, Bufei Jianpi granules: Implication for COPD drug …Pubmed:35121390
  • IL4 stimulated macrophages promote axon regeneration after peripheral nerve injury by secreting uPA to stimulate uPAR upregulated in injured axonsPubmed:35536429

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