High Sensitive ELISA Kit for Matrix Metalloproteinase 1 (MMP1)

CLGN; CLG1; Collagenase; Interstitial Collagenase; Vertebrate Collagenase; Fibroblast Collagenase

Specificity

This assay has high sensitivity and excellent specificity for detection of High Sensitive Matrix Metalloproteinase 1 (MMP1).
No significant cross-reactivity or interference between High Sensitive Matrix Metalloproteinase 1 (MMP1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant High Sensitive Matrix Metalloproteinase 1 (MMP1) and the recovery rates were calculated by comparing the measured value to the expected amount of High Sensitive Matrix Metalloproteinase 1 (MMP1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-99 83
heparin plasma(n=5) 87-94 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Matrix Metalloproteinase 1 (MMP1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Matrix Metalloproteinase 1 (MMP1) 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 High Sensitive Matrix Metalloproteinase 1 (MMP1) 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) 78-104% 84-97% 91-101% 78-96%
heparin plasma(n=5) 85-99% 88-104% 91-105% 81-99%

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.

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Magazine Citations
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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
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
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Int Wound J. From varices to venous ulceration: the story of chronic venous disease described by metalloproteinases. Pubmed:26991748
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Molecular Medicine Reports Icariin inhibits MMP‑1, MMP‑3 and MMP‑13 expression through MAPK pathways in IL‑1β‑stimulated SW1353 chondrosarcoma cells pubmed:28447732
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Journal of Applied Animal Research Hypoxia enhances secretion of matrix metalloproteinases in bovine dermal fibroblasts:an in vitro approach to bovine digital dermatitis 10.108:09712119.2017.1335208
Analytical Methods SPRI biosensors for quantitative determination of matrix metalloproteinase-2 DOI:10.1039/C7AY00786H
International Wound Journal From varices to venous ulceration: the story of chronic venous disease described by metalloproteinases pubmed:26991748
European Heart Journal Biomarker‐based phenotyping of myocardial fibrosis identifies patients with heart failure with preserved ejection fraction resistant to the beneficial effects of … Pubmed:29709099
Journal of Agricultural and Food Chemistry Hawthorn polyphenol extract inhibits UVB-induced skin photoaging by regulating MMP expression and type I procollagen production in mice Pubmed:30032605
Oral Diseases  TIMP‐1 association with collagen type I overproduction in hereditary gingival fibromatosis Pubmed:29989318
Journal of Applied Animal Research  Hypoxia enhances secretion of matrix metalloproteinases in bovine dermal fibroblasts: an approach to bovine digital dermatitis 10.1080:09712119.2017.1335208
PLoS One Co-culture of osteochondral explants and synovial membrane as in vitro model for osteoarthritis Pubmed: 30939166
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