ELISA Kit for Collagen Type II (COL2) Homo sapiens (Human) Sandwich ELISA

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  • ELISA Kit for Collagen Type II (COL2) Packages (Simulation)
  • ELISA Kit for Collagen Type II (COL2) Packages (Simulation)
  • ELISA Kit for Collagen Type II (COL2) Results demonstration
  • SEA572Hu.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) and the recovery rates were calculated by comparing the measured value to the expected amount of Collagen Type II (COL2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-99 86
EDTA plasma(n=5) 91-105 97
heparin plasma(n=5) 78-89 81

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Collagen Type II (COL2) 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) 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) 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) 82-101% 96-105% 93-101% 84-104%
EDTA plasma(n=5) 95-105% 85-103% 80-102% 97-104%
heparin plasma(n=5) 96-105% 80-101% 90-97% 81-95%

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.

ELISA Kit for Collagen Type II (COL2)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Collagen Type II (COL2). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Collagen Type II (COL2). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Collagen Type II (COL2), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Collagen Type II (COL2) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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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