Multiplex Assay Kit for Matrix Metalloproteinase 13 (MMP13) ,etc. by FLIA (Flow Luminescence Immunoassay)

CLG3; Collagenase 3

(Note: Up to 8-plex in one testing reaction)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 87-101 95
EDTA plasma(n=5) 84-105 97
heparin plasma(n=5) 80-89 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Matrix Metalloproteinase 13 (MMP13) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 83-97% 94-105% 95-104% 94-101%
EDTA plasma(n=5) 82-104% 91-105% 85-92% 96-104%
heparin plasma(n=5) 81-97% 79-95% 79-95% 83-102%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:MMP13) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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
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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
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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
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Molecular Medicine Reports Effects of GGCX overexpression on anterior cruciate ligament transection-induced osteoarthritis in rabbits Pubmed:29257344
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Nature Communications Vulnerability of progeroid smooth muscle cells to biomechanical forces is mediated by MMP13 Pubmed: 32807790
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