Multiplex Assay Kit for Matrix Metalloproteinase 8 (MMP8) ,etc. by CBA (Cytometric Bead Array)

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) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Matrix Metalloproteinase 8 (MMP8) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 87-94 90
EDTA plasma(n=5) 92-101 96
heparin plasma(n=5) 80-96 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Matrix Metalloproteinase 8 (MMP8) ,etc. by CBA (Cytometric Bead Array) 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) ,etc. by CBA (Cytometric Bead Array) 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) ,etc. by CBA (Cytometric Bead Array) 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) 85-97% 79-97% 86-104% 89-97%
EDTA plasma(n=5) 92-101% 79-101% 98-105% 80-99%
heparin plasma(n=5) 98-105% 85-99% 85-97% 98-105%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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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
International Wound Journal From varices to venous ulceration: the story of chronic venous disease described by metalloproteinases pubmed:26991748
Medicine (Baltimore) Rapid detection of urinary soluble intercellular adhesion molecule-1 for determination of lupus nephritis activity Pubmed:29953010
Scientific Reports Atherosclerotic plaque vulnerability is increased in mouse model of lupus Pubmed: 33110193
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