Properties
Assay type | Sandwich-ELISA |
Format | 96T/48T |
Assay time | 3.5h |
Detection range | 0.78-50 ng/mL |
Sensitivity | 0.47 ng/mL |
Sample type &Sample volume | serum, plasma and other biological fluids; 100μL |
Specificity | This kit recognizes Human MMP-2 in samples. No significant cross-reactivity or interference between Human MMP-2 and analogues was observed. |
Reproducibility | Both intra-CV and inter-CV are < 10%. |
Application | This ELISA kit applies to the in vitro quantitative determination of Human MMP-2 concentrations in serum, plasma and other biological fluids. |
Test Principle
This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human MMP-2. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human MMP-2 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human MMP-2, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human MMP-2. You can calculate the concentration of Human MMP-2 in the samples by comparing the OD of the samples to the standard curve.
Kit components & Storage
An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
Item | Specifications | Storage |
---|---|---|
Micro ELISA Plate(Dismountable) | 96T: 8 wells ×12 strips 48T: 8 wells ×6 strips |
-20℃, 6 months |
Reference Standard | 96T: 2 vials 48T: 1 vial |
|
Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL 48T: 1 vial, 60 μL |
|
Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL 48T: 1 vial, 60 μL |
-20℃(Protect from light), 6 months |
Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
HRP Conjugate Diluent | 1 vial, 14 mL | |
Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
Substrate Reagent | 1 vial, 10 mL | 2-8℃(Protect from light) |
Stop Solution | 1 vial, 10 mL | 2-8°C |
Plate Sealer | 5 pieces | |
Manual | 1 copy | |
Certificate of Analysis | 1 copy |
Technical Data
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human MMP-2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human MMP-2 were tested on 3 different plates, 20 replicates in each plate.
Intra-assay Precision | Inter-assay Precision | |||||
---|---|---|---|---|---|---|
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 20 | 20 | 20 |
Mean (ng/mL) | 2.40 | 5.30 | 21.80 | 2.40 | 5.10 | 20.80 |
Standard deviation | 0.10 | 0.30 | 0.90 | 0.20 | 0.20 | 0.70 |
CV (%) | 4.17 | 5.66 | 4.13 | 8.33 | 3.92 | 3.37 |
Recovery
The recovery of Human MMP-2 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type | Range (%) | Average Recovery (%) |
---|---|---|
Serum(n=8) | 86-100 | 92 |
EDTA plasma (n=8) | 87-99 | 92 |
Cell culture media (n=8) | 88-100 | 93 |