ELISA Kit for A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4) Homo sapiens (Human) Sandwich ELISA

ADMP-1; Aggrecanase-1

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  • ELISA Kit for A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4) Packages (Simulation)
  • ELISA Kit for A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4) Packages (Simulation)
  • ELISA Kit for A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4) Results demonstration
  • SEK204Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4) and the recovery rates were calculated by comparing the measured value to the expected amount of A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 96-103 101
EDTA plasma(n=5) 97-104 101
heparin plasma(n=5) 80-97 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4) 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 A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4) 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) 90-104% 95-104% 97-105% 88-97%
EDTA plasma(n=5) 96-103% 78-91% 89-96% 95-105%
heparin plasma(n=5) 78-95% 83-101% 84-95% 78-99%

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 A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4)

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 A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4). 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 A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4), 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 A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • ADAMTS4 level in patients with stable coronary artery disease and acute coronary syndromesScienceDirect: S0753332209002017
  • A Combinatorial Relative Mass Value Evaluation of Endogenous Bioactive Proteins in Three-Dimensional Cultured Nucleus Pulposus Cells of Herniated Intervertebral Discs: Identification of Potential Target Proteins for Gene Therapeutic ApproachesPlosone: Source
  • A novel association between TGFb1 and ADAMTS4 in coronary artery disease: A new potential mechanism in the progression of atherosclerosis and diabetes.Pubmed:25592103
  • Lack of anti-inflammatory and anti-catabolic effects on basal inflamed osteoarthritic chondrocytes or synoviocytes by adipose stem cell-conditioned mediumPubMed: 26521751
  • Parathyroid hormone(1-34) exhibits more comprehensive effects than celecoxib in cartilage metabolism and maintaining subchondral bone micro-architecture in meniscectomized Cavia (Guinea pig )inea pigsPubmed:26802547
  • VIP and CRF reduce ADAMTS expression and function in osteoarthritis synovial fibroblastsPubmed:26818776
  • From varices to venous ulceration: the story of chronic venous disease described by metalloproteinases.Pubmed:26991748
  • Healthy and Osteoarthritic Synovial Fibroblasts Produce a Disintegrin and Metalloproteinase with Thrombospondin Motifs 4, 5, 7, and 12: Induction by IL-1β and Fibronectin and Contribution to Cartilage Damage.pubmed:27449198
  • From varices to venous ulceration: the story of chronic venous disease described by metalloproteinasespubmed:26991748
  • Diagnostic utility of fluorogenic peptide-conjugated Au nanoparticle probe corroborated by rabbit model of mild cartilage injury and panel of osteoarthritic …Pubmed: 30210670
  • The Effects of TNF-α Inhibition on the Metabolism of Cartilage: Relationship between KS, HA, HAPLN1 and ADAMTS4, ADAMTS5, TOS and TGF-β1 Plasma …Pubmed:35407621

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