Human COMP Quantikine ELISA Kit

(1 Review)
  
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
  • Sensitivity
    0.036 ng/mL
  • Assay Range
    0.2 - 10 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant human COMP
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC66, Quantikine Immunoassay Control Set 702 for Human COMP - Please Inquire
Product Summary
The Quantikine Human COMP immunoassay is a 4.5 hour solid phase ELISA designed to measure human COMP in cell culture supernates, cell lysates, serum, and plasma. It contains NS0-expressed recombinant human COMP and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human COMP showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring COMP.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean0.92.926.111.053.076.26
Standard Deviation0.0280.1120.2250.0450.1460.3
CV%2.13.83.74.34.84.8

Recovery

The recovery of COMP spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 103 95-111
Cell Lysates (n=1) 102 98-108
EDTA Plasma (n=4) 103 86-114
Heparin Plasma (n=4) 99 88-112
Serum (n=4) 102 86-114
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human COMP were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human COMP Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: COMP/Thrombospondin-5
COMP (Cartilage Oligomeric Matrix Protein), also known as Thrombospondin-5, is a multidomain calcium binding protein that associates with other extracellular matrix molecules. COMP associates into disulfide-linked homopentamers with a central hub and peripheral globular domains connected by flexible strands. An axial pore is formed by the coiled coil assembly and binds vitamin D3 which is involved in bone and cartilage metabolism. An RGD sequence in the third TSP type-3 repeat mediates chondrocyte attachment via Integrins a5b1 or aVb3. COMP is upregulated in rheumatoid arthritis and osteoarthritis, hepatocellular carcinomas, chronic pancreatitis, and pancreatic carcinomas. Several mutations in COMP are associated with skeletal dysplasias.
    • Long Name
      Cartilage Oligomeric Matrix Protein
    • Entrez Gene IDs
      1311 (Human); 12845 (Mouse);
    • Alternate Names
      cartilage oligomeric matrix protein (pseudoachondroplasia, epiphyseal dysplasia1, multiple); cartilage oligomeric matrix protein; cartilage oligomeric matrix protein(pseudoachondroplasia, epiphyseal dysplasia1, multiple); COMP; EDM1; EPD1; MED; MEDMGC131819; MGC149768; PSACH; pseudoachondroplasia (epiphyseal dysplasia 1, multiple); THBS5; Thrombospondin5; Thrombospondin-5; TSP5;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 702 for Human COMP

    Ctrl QC66
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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    Cell Lysis Buffer 1 (1 x 21 mL)

    ELISA 890713 1
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    Average Rating: 4 (Based on 1 review)

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