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Human MMP-9 (Matrix Metalloproteinase 9) CLIA Kit

  • Cat.No.:E-CL-H0936

  • Reactivity: Human

To Purchase E-CL-H0936

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $682
Qty:

Product Details

Properties

Assay type Sandwich-CLIA
Format 96T/48T
Assay time 3.5h
Detection range 62.50-4000 pg/mL
Sensitivity 37.50 pg/mL
Sample type &Sample volume serum, plasma and other biological fluids; 100μL
Specificity This kit recognizes Human MMP-9 in samples. No significant cross-reactivity or interference between Human MMP-9 and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 15%.
Application This CLIA kit applies to the in vitro quantitative determination of Human MMP-9 concentrations in serum, plasma and other biological fluids.

Test Principle

This CLIA kit uses the Sandwich-CLIA principle. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human MMP-9. Standards or samples are added to the micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human MMP-9 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-9, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human MMP-9. You can calculate the concentration of Human MMP-9 in the samples by comparing the RLU value 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.
ItemSpecificationsStorage
Micro CLIA 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 A 1 vial, 5 mL 2-8℃(Protect from light)
Substrate Reagent B 1 vial, 5 mL
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-9 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-9 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 (pg/mL) 220.89 625.04 1419.98 223.42 687.02 1340.57
Standard deviation 20.76 47.38 127.09 23.59 74.54 142.37
CV (%) 9.40 7.58 8.95 10.56 10.85 10.62

Recovery

The recovery of Human MMP-9 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) 98-112 104
EDTA plasma(n=8) 92-109 99
Cell culture media(n=8) 102-116 107

Linearity

Samples were spiked with high concentrations of Human MMP-9 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

SynonymsMMP9, CLG4B,Gelatinase B, GELB, MANDP2, 92kDa Type IV Collagenase, 92 KDa Gelatinase

Assay Procedures

elisa assay procedure 1

1.Add 100 μL of standard or sample to each well. Incubate for 90 min at 37℃.

elisa assay procedure 2

2.Remove the liquid.

elisa assay procedure 3

3.Add 100 μL Biotinylated Detection Ab. Incubate 1 hour at 37℃. Aspirate and wash 3 times.

elisa assay procedure 4

4.Add 100 μL HRP Conjugate. Incubate 30 min at 37℃. Aspirate and wash 5 times.

elisa assay procedure 5

5.Add 100 μL Substrate Mixture Solution. Incubate for 5 min at 37℃.

elisa assay procedure 6

6.Fluorescence appeared. Measure the RLU value with the Chemiluminescence immunoassay analyzer. Calculation of the results.

Citations

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