CLIA Kit for Matrix Metalloproteinase 13 (MMP13)

CLG3; Collagenase 3

Specificity

This assay has high sensitivity and excellent specificity for detection of Matrix Metalloproteinase 13 (MMP13).
No significant cross-reactivity or interference between Matrix Metalloproteinase 13 (MMP13) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Matrix Metalloproteinase 13 (MMP13) and the recovery rates were calculated by comparing the measured value to the expected amount of Matrix Metalloproteinase 13 (MMP13) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-104 101
heparin plasma(n=5) 95-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Matrix Metalloproteinase 13 (MMP13) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Matrix Metalloproteinase 13 (MMP13) 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 13 (MMP13) 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) 85-99% 90-101% 86-101% 98-105%
heparin plasma(n=5) 78-93% 90-97% 82-101% 89-97%

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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

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Magazine Citations
Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease Hyaluronan reduces inflammation in experimental arthritis by modulating TLR-2 and TLR-4 cartilage expression ScienceDirect: S0925443911001323
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FEBS Journal Adenosine A2a Receptor Activation and Hyaluronan Fragment Inhibition Reduce Inflammation in Mouse Articular Chondrocytes Stimulated with Il-1beta Wiley: source
Arthritis & Rheumatism Interruption of glycosphingolipid synthesis enhances osteoarthritis development in mice Wiley: source
Gene Hyaluronan in part mediates IL-1beta-induced inflammation in mouse chondrocytes by up-regulating CD44 receptors ScienceDirect: S0378111911007347
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
Arthritis Rheumatol Identification of Alpha 2 Macroglobulin (A2M) as a master inhibitor of cartilage degrading factors that attenuates post-traumatic osteoarthritis progression. Pubmed: 24578232
Arthritis & Rheumatology Identification of Alpha 2 Macroglobulin (A2M) as a master inhibitor of cartilage degrading factors that attenuates post-traumatic osteoarthritis progression Onlinelibrary: art.38576
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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Osteoarthritis and Cartilage Depletion of gangliosides enhances cartilage degradation in mice Sciencedirect:S1063458413010364
Biol Res The role of MCP-1-CCR2 ligand-receptor axis in chondrocyte degradation and disease progress in knee osteoarthritis PubMed: 26578310
PLoS One Interleukin-1 Receptor Antagonist Has a Novel Function in the Regulation of Matrix Metalloproteinase-13 Expression PubMed: 26474296
Cell Mol Immunol  Toxoplasma gondii GRA15II effector-induced M1 cells ameliorate liver fibrosis in mice infected with Schistosomiasis japonica Pubmed:27157496
Scientific Reports Chondrogenic Potential of Peripheral Blood Derived Mesenchymal Stem Cells Seeded on Demineralized Cancellous Bone Scaffolds pubmed:27821864
International Journal of Contemporary Medical Research Matrix Metalloproteinase-13 (MMP-13): A Novel Tumor Marker for Diagnosis of Breast Carcinoma uploads:7
Scientific Reports Olive and grape seed extract prevents post-traumatic osteoarthritis damages and exhibits in vitro anti IL-1β activities before and after oral consumption. pubmed:27640363
Photodermatol Photoimmunol Photomed. Protective effects of Aloe sterols against UVB‐induced photoaging in hairless mice pubmed:27995657
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
bmc musculoskeletal disorders Strontium ranelate causes osteophytes overgrowth in a model of early phase osteoarthritis. pubmed:28187731
Journal of Cellular Physiology Increased Chondrogenic Potential of Mesenchymal Cells From Adipose Tissue Versus Bone Marrow‐Derived Cells in Osteoarthritic In Vitro Models pubmed:27739057
BMC Musculoskeletal Disorders Establishment of rat ankle post-traumatic osteoarthritis model induced by malleolus fracture pubmed:29149841
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
Alexandria Journal of Medicine L-Carnitine ameliorates knee lesions in mono-iodoacetate induced osteoarthritis in rats 10.1016:j.ajme.2016.03.002
National Journal of Integrated Research in Medicine Matrix Metalloproteinase-13 Can Be Used As an Independent Diagnostic and Prognostic Marker for Breast Carcinoma. AN=123116960
Molecular Medicine Reports Effects of GGCX overexpression on anterior cruciate ligament transection-induced osteoarthritis in rabbits Pubmed:29257344
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
Osteoarthritis Cartilage Coordinated existence of multiple gangliosides is required for cartilage metabolism Pubmed: 30471358
PLoS One Co-culture of osteochondral explants and synovial membrane as in vitro model for osteoarthritis Pubmed: 30939166
Pathobiology Suppressed Expression but Not Activity of Collagenases MMP-1 and MMP-13 in Human Renal Carcinoma Pubmed: 31129674
Bone DAla2GIP antagonizes H2O2-induced chondrocyte apoptosis and inflammatory factor secretion Pubmed: 31283994
Food & Function A dietary polysaccharide from Eucheuma cottonii downregulates proinflammatory cytokines and ameliorates osteoarthritis-associated cartilage degradation in obese …
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Nature Communications Vulnerability of progeroid smooth muscle cells to biomechanical forces is mediated by MMP13 Pubmed: 32807790
Molecular Medicine Reports MicroRNA-1 promotes cartilage matrix synthesis and regulates chondrocyte differentiation via post-transcriptional suppression of Ihh expression Pubmed: 32705199
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