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ELISA Kit for Matrix Metalloproteinase 8 (MMP8)

CLG2; HNC; PMNL-CL; Collagenase 2; Neutrophil Collagenase; PMNL collagenase

Specificity

This assay has high sensitivity and excellent specificity for detection of Matrix Metalloproteinase 8 (MMP8).
No significant cross-reactivity or interference between Matrix Metalloproteinase 8 (MMP8) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Matrix Metalloproteinase 8 (MMP8) and the recovery rates were calculated by comparing the measured value to the expected amount of Matrix Metalloproteinase 8 (MMP8) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 101
heparin plasma(n=5) 98-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Matrix Metalloproteinase 8 (MMP8) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Matrix Metalloproteinase 8 (MMP8) 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 Matrix Metalloproteinase 8 (MMP8) 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) 95-103% 79-99% 91-98% 96-103%
heparin plasma(n=5) 80-96% 84-93% 98-105% 94-105%

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.

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Magazine Citations
PLoS ONE 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 Approaches Plosone: Source
Journal of Periodontology Effects of Photodynamic Therapy on the Clinical and Gingival Crevicular Fluid Inflammatory Biomarkers in Chronic Periodontitis: A Split-Mouth Randomized Clinical Trial Joponline: 130464
Thromb Haemost. Analysis of the expression of nine secreted matrix metalloproteinases and their endogenous inhibitors in the brain of mice subjected to ischaemic stroke Pubmed:24671655
An Experimental Model for Inducing Periodontal Pathology in Rat: Histopathological and Enzymatic Aspects Usamvcluj:Source
Journal of Molecular and Cellular Cardiology Treatment with anti-RANKL antibody reduces infarct size and attenuates dysfunction impacting on neutrophil-mediated injury Pubmed:27056420
J Periodontal Implant Sci Assessment of MMP-1, MMP-8 and TIMP-2 in experimental periodontitis treated with kaempferol pmc:PMC4848383
Int Wound J. From varices to venous ulceration: the story of chronic venous disease described by metalloproteinases. Pubmed:26991748
Eur J Clin Invest.  Alamandine abrogates neutrophil degranulation in atherosclerotic mice. pubmed:27930810
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