Multiplex Assay Kit for Collagen Type II (COL2) ,etc. by FLIA (Flow Luminescence Immunoassay) Rattus norvegicus (Rat) Multiplex ELISA

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

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  • Multiplex Assay Kit for Collagen Type II (COL2) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Collagen Type II (COL2) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Collagen Type II (COL2) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA572Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Collagen Type II (COL2) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Collagen Type II (COL2) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-95 81
EDTA plasma(n=5) 92-105 102
heparin plasma(n=5) 98-105 101
sodium citrate plasma(n=5) 98-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Collagen Type II (COL2) ,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 Collagen Type II (COL2) ,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 Collagen Type II (COL2) ,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) 82-90% 88-104% 96-104% 84-93%
EDTA plasma(n=5) 80-97% 98-105% 79-92% 96-103%
heparin plasma(n=5) 85-104% 85-94% 86-99% 98-105%
sodium citrate plasma(n=5) 85-99% 80-96% 79-96% 98-105%

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#:COL2) 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 Collagen Type II (COL2) ,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.

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Citations

  • Elevated PLGF contributes to small-cell lung cancer brain metastasisPubMed: 22797069
  • Acidic pH conditions mimicking degenerative intervertebral discs impair the survival and biological behavior of human adipose-derived mesenchymal stem cellsRsmjournals: Source
  • 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
  • Protein synthesis and secretion in human mesenchymal cells derived from bone marrow, adipose tissue and Wharton's jellyPubmed: 24739658
  • No effects of Hyperosmolar Culture medium on Tissue Regeneration by Human Degenerated Nucleus Pulposus Cells despite Upregulation Extracellular Matrix Genes.Pubmed:25856264
  • Mesenchymal stromal cell proliferation, gene expression and protein production in human platelet-rich plasma-supplemented mediaPubmed:Pmc4130592
  • Co-electrospun gelatin-poly (l-lactic acid) scaffolds: modulation of mechanical properties and chondrocyte response as a function of compositionPubmed:24433895
  • 可拋棄式退化性關節炎電化學式尿液免疫感測器之研究 Electrochemical Immunosensor Detection of Urinary Biomaker for Osteoarthritis Diagnosis Publication: U0074-2807201500521300
  • Expression and significance of telomerase in the nucleus pulposus tissues of degenerative lumbar discsPubMed: 26623021
  • Highly Porous Gelatin Reinforced 3D Scaffolds for Articular Cartilage RegenerationPubMed: 25787871
  • The role of MCP-1-CCR2 ligand-receptor axis in chondrocyte degradation and disease progress in knee osteoarthritisPubMed: 26578310
  • Effects of low oxygen tension on gene profile of soluble growth factors in co‐cultured adipose‐derived stromal cells and chondrocytesPubmed:27090063
  • 白细胞介素10 上调表达对家兔慢性细菌性鼻窦炎黏膜创伤修复的影响article:11593
  • An innovative co-axial system to electrospin in situ crosslinked gelatin nanofibers.Pubmed:26987305
  • Chondrogenic Potential of Peripheral Blood Derived Mesenchymal Stem Cells Seeded on Demineralized Cancellous Bone Scaffoldspubmed:27821864
  • Self-complementary adeno-associated virus serotype 6 mediated knockdown of ADAMTS4 induces long-term and effective enhancement of aggrecan in degenerative human nucleus pulposus cells: A new therapeutic approach for intervertebral disc disorders.pubmed:28207788
  • Microsphere-Based Hierarchically Juxtapositioned Biphasic Scaffolds Prepared from Poly(Lactic-co-Glycolic Acid) and Nanohydroxyapatite for Osteochondral Tissue Engineering2073-4360:8
  • Knockout of Apolipoprotein E in rabbit promotes premature intervertebral disc degeneration: A new in vivo model for therapeutic approaches of spinal disc disorders10.1371:journal.pone.0187564
  • Self-complementary adeno-associated virus serotype 6 mediated knockdown of ADAMTS4 induces long-term and effective enhancement of aggrecan in degenerative human nucleus pulposus cells: A new therapeutic approach for intervertebral disc disorders10.1371:journal.pone.0172181
  • Platelet-rich concentrate in serum-free medium enhances cartilage-specific extracellular matrix synthesis and reduces chondrocyte hypertrophy of human mesenchymal stromal cells encapsulated in alginate.pubmed:29090639
  • Effects of GGCX overexpression on anterior cruciate ligament transection-induced osteoarthritis in rabbitsPubmed:29257344
  • Scaffolds modified with graphene as future implants for nasal cartilage
  • A Novel Method to Optimize Autologous Adipose Tissue Recovery with Extracellular Matrix Preservation

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