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 Matrix Metalloproteinase 1 (MMP1).
No significant cross-reactivity or interference between Matrix Metalloproteinase 1 (MMP1) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 92-105 97
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 1 (MMP1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level 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 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) 87-95% 99-105% 90-98% 78-96%
heparin plasma(n=5) 86-94% 81-99% 83-101% 82-90%

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
Parasitology Research Therapeutic effect of alpha lipoic acid combined with praziquantel on liver fibrosis induced by Schistosoma mansoni challenged mice SpringerLink: j33n80517577535k
Infection and Immunity Bovine Respiratory Syncytial Virus and Histophilus somni Interaction at the Alveolar Barrier Pubmed: 23649093
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
The Journal of Pathology Mycobacterium tuberculosis dysregulates MMP/TIMP balance to drive rapid cavitation and unrestrained bacterial proliferation Pubmed:25186281
Connective Tissue Research Metabolic and cytoprotective effects of in vivo peri-patellar hyaluronic acid injections in cultured tenocytes Pubmed:25333747
J Immunol Heme Oxygenase-1 Regulation of Matrix Metalloproteinase-1 Expression Underlies Distinct Disease Profiles in Tuberculosis PubMed: 26268658
Cellular & Molecular Immunology Effect of ebosin on modulating interleukin-1&bgr;-induced inflammatory responses in rat fibroblast-like synoviocytes PubMed: 25938977
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
Cellular & Molecular Immunology Effect of ebosin on modulating interleukin-1β-induced inflammatory responses in rat fibroblast-like synoviocytes pubmed:25938977
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
Analytical Methods The development of a matrix metalloproteinase-1 biosensor based on the surface plasmon resonance imaging technique c6ay01856d
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