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

CLG3; Collagenase 3

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

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  • Multiplex Assay Kit for Matrix Metalloproteinase 13 (MMP13) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Matrix Metalloproteinase 13 (MMP13) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Matrix Metalloproteinase 13 (MMP13) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA099Ga.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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) 95-102 98
EDTA plasma(n=5) 97-105 101
heparin plasma(n=5) 79-102 83

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) 79-105% 80-101% 92-101% 85-95%
EDTA plasma(n=5) 85-104% 97-104% 82-89% 78-90%
heparin plasma(n=5) 87-96% 80-104% 79-89% 83-93%

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.

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

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

Citations

  • Hyaluronan reduces inflammation in experimental arthritis by modulating TLR-2 and TLR-4 cartilage expressionScienceDirect: S0925443911001323
  • Protective effects of curcumin, α-lipoic acid, and N-acetylcysteine against carbon tetrachloride-induced liver fibrosis in ratsSpringerLink: h875w73171w2147w
  • Adenosine A2a Receptor Activation and Hyaluronan Fragment Inhibition Reduce Inflammation in Mouse Articular Chondrocytes Stimulated with Il-1betaWiley: source
  • Interruption of glycosphingolipid synthesis enhances osteoarthritis development in miceWiley: source
  • Hyaluronan in part mediates IL-1beta-induced inflammation in mouse chondrocytes by up-regulating CD44 receptorsScienceDirect: S0378111911007347
  • 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 ApproachesPlosone: Source
  • Identification of Alpha 2 Macroglobulin (A2M) as a master inhibitor of cartilage degrading factors that attenuates post-traumatic osteoarthritis progression.Pubmed: 24578232
  • Identification of Alpha 2 Macroglobulin (A2M) as a master inhibitor of cartilage degrading factors that attenuates post-traumatic osteoarthritis progressionOnlinelibrary: art.38576
  • Analysis of the expression of nine secreted matrix metalloproteinases and their endogenous inhibitors in the brain of mice subjected to ischaemic strokePubmed:24671655
  • Identification of α2-Macroglobulin as a Master Inhibitor of Cartilage-Degrading Factors That Attenuates the Progression of Posttraumatic OsteoarthritisPubmed:24578232
  • Systemic and Flap Inflammatory Response Associates with Thrombosis in Flap Venous CrisisPubmed:25448261
  • Inverse Regulation of Early and Late Chondrogenic Differentiation by Oxygen Tension Provides Cues for Stem Cell-Based Cartilage Tissue EngineeringPubmed:25632940
  • Depletion of gangliosides enhances cartilage degradation in miceSciencedirect:S1063458413010364
  • The role of MCP-1-CCR2 ligand-receptor axis in chondrocyte degradation and disease progress in knee osteoarthritisPubMed: 26578310
  • Interleukin-1 Receptor Antagonist Has a Novel Function in the Regulation of Matrix Metalloproteinase-13 ExpressionPubMed: 26474296
  • Toxoplasma gondii GRA15II effector-induced M1 cells ameliorate liver fibrosis in mice infected with Schistosomiasis japonicaPubmed:27157496
  • Chondrogenic Potential of Peripheral Blood Derived Mesenchymal Stem Cells Seeded on Demineralized Cancellous Bone Scaffoldspubmed:27821864
  • Matrix Metalloproteinase-13 (MMP-13): A Novel Tumor Marker for Diagnosis of Breast Carcinomauploads:7
  • 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
  • Protective effects of Aloe sterols against UVB‐induced photoaging in hairless micepubmed:27995657
  • Icariin inhibits MMP‑1, MMP‑3 and MMP‑13 expression through MAPK pathways in IL‑1β‑stimulated SW1353 chondrosarcoma cellspubmed:28447732
  • Strontium ranelate causes osteophytes overgrowth in a model of early phase osteoarthritis.pubmed:28187731
  • Increased Chondrogenic Potential of Mesenchymal Cells From Adipose Tissue Versus Bone Marrow‐Derived Cells in Osteoarthritic In Vitro Modelspubmed:27739057
  • Establishment of rat ankle post-traumatic osteoarthritis model induced by malleolus fracturepubmed:29149841
  • Hypoxia enhances secretion of matrix metalloproteinases in bovine dermal fibroblasts:an in vitro approach to bovine digital dermatitis 10.108:09712119.2017.1335208
  • L-Carnitine ameliorates knee lesions in mono-iodoacetate induced osteoarthritis in rats10.1016:j.ajme.2016.03.002
  • Matrix Metalloproteinase-13 Can Be Used As an Independent Diagnostic and Prognostic Marker for Breast Carcinoma.AN=123116960
  • Effects of GGCX overexpression on anterior cruciate ligament transection-induced osteoarthritis in rabbitsPubmed:29257344
  • Hypoxia enhances secretion of matrix metalloproteinases in bovine dermal fibroblasts: an approach to bovine digital dermatitis10.1080:09712119.2017.1335208
  • Coordinated existence of multiple gangliosides is required for cartilage metabolismPubmed: 30471358
  • Co-culture of osteochondral explants and synovial membrane as in vitro model for osteoarthritisPubmed: 30939166
  • Suppressed Expression but Not Activity of Collagenases MMP-1 and MMP-13 in Human Renal CarcinomaPubmed: 31129674
  • DAla2GIP antagonizes H2O2-induced chondrocyte apoptosis and inflammatory factor secretionPubmed: 31283994
  • A dietary polysaccharide from Eucheuma cottonii downregulates proinflammatory cytokines and ameliorates osteoarthritis-associated cartilage degradation in obese …
  • Teneligliptin inhibits IL‐1β‐induced degradation of extracellular matrix in human chondrocytesPubmed: 32162384
  • Effects of tetrahedral framework nucleic acid/wogonin complexes on osteoarthritisPubmed: 32047705
  • Matrix metallopeptidase expression and modulation by transforming growth factor-β1 in equine endometrosisPubmed: 31980722
  • Chondroprotective Properties of Human-Enriched Serum Following Polyphenol Extract Absorption: Results from an Exploratory Clinical TrialPubmed: 31888255
  • Vulnerability of progeroid smooth muscle cells to biomechanical forces is mediated by MMP13Pubmed: 32807790
  • MicroRNA-1 promotes cartilage matrix synthesis and regulates chondrocyte differentiation via post-transcriptional suppression of Ihh expressionPubmed: 32705199
  • Experimental Observation of the Sequence of Tibial Plateau Chondrocyte and Matrix Degeneration in Spontaneous Osteoarthritis in Guinea Pigs
  • Hyaluronic Acid versus Alpha-2-Macroglobulin Intra-Articular Injections for Amelioration of Knee Posttraumatic Osteoarthritis: A Rat Model
  • Loss of autophagy in tibial plateau chondrocytes causes increased apoptosis of chondrocytes in spontaneous osteoarthritis of guinea pigs
  • Undenatured type II collagen prevents and treats the osteoarthritis and motor function degradation in T2DM patients and db/db mice33890588
  • Metabolic and Anti-Inflammatory Protective Properties of Human Enriched Serum Following Artichoke Leaf Extract Absorption: Results from an Innovative Ex Vivo …34444810
  • Liraglutide, a glucagon-like peptide 1 receptor agonist, exerts analgesic, anti-inflammatory and anti-degradative actions in osteoarthritisPubmed:35091584

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