High Sensitive ELISA Kit for Matrix Metalloproteinase 13 (MMP13)

CLG3; Collagenase 3

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 82-96 90
heparin plasma(n=5) 82-89 85

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive 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 High Sensitive 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 High Sensitive 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) 86-104% 84-91% 83-92% 88-102%
heparin plasma(n=5) 79-101% 89-102% 99-105% 96-103%

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
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
Journal of Physiology and Biochemistry Protective effects of curcumin, α-lipoic acid, and N-acetylcysteine against carbon tetrachloride-induced liver fibrosis in rats SpringerLink: h875w73171w2147w
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
Arthritis &amp; Rheumatol Identification of α2-Macroglobulin as a Master Inhibitor of Cartilage-Degrading Factors That Attenuates the Progression of Posttraumatic Osteoarthritis Pubmed:24578232
Inflammation.? Systemic and Flap Inflammatory Response Associates with Thrombosis in Flap Venous Crisis Pubmed:25448261
Cellular Physiology and Biochemistry Inverse Regulation of Early and Late Chondrogenic Differentiation by Oxygen Tension Provides Cues for Stem Cell-Based Cartilage Tissue Engineering Pubmed:25632940
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
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