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ELISA Kit for Cartilage Oligomeric Matrix Protein (COMP)

MED; EDM1; EPD1; PSACH; THBS5; TSP-5; Pseudoachondroplasia,Epiphyseal Dysplasia 1,Multiple; Thrombospondin 5

Specificity

This assay has high sensitivity and excellent specificity for detection of Cartilage Oligomeric Matrix Protein (COMP).
No significant cross-reactivity or interference between Cartilage Oligomeric Matrix Protein (COMP) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Cartilage Oligomeric Matrix Protein (COMP) and the recovery rates were calculated by comparing the measured value to the expected amount of Cartilage Oligomeric Matrix Protein (COMP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-98 83
EDTA plasma(n=5) 78-98 94
heparin plasma(n=5) 79-103 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cartilage Oligomeric Matrix Protein (COMP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cartilage Oligomeric Matrix Protein (COMP) 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 Cartilage Oligomeric Matrix Protein (COMP) 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) 83-101% 94-101% 93-101% 95-103%
EDTA plasma(n=5) 97-104% 81-102% 88-102% 97-105%
heparin plasma(n=5) 94-105% 80-88% 98-105% 81-104%

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.

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Magazine Citations
Chinese Medical Journal Genetic analysis and serum level of cartilage oligomeric matrix protein in patients with pseudoachondroplasia CMJ: 201082337960590
BioMed Research International Decreased Plasma COMP and Increased Plasma CTX-II Levels in a Chinese Pseudoachondroplasia Family with Novel COMP Mutation pubmed:29104872
Molecular Medicine Reports Mori Folium water extract alleviates articular cartilage damages and inflammatory responses in monosodium iodoacetate‑induced osteoarthritis rats pubmed:29067461
Archives of Rheumatology Cartilage Oligomeric Matrix Protein Levels in Synovial Fluid in Patients With Primary Knee Osteoarthritis And Healthy Controls: A Preliminary Comparative Analysis With Serum Cartilage Oligomeric Matrix Protein doi: 10.5606/ArchRheumatol.2017.6220
Genetic Testing and Molecular Biomarkers Cartilage Oligomeric Matrix Protein Levels in Type 2 Diabetes Associated with Primary Knee Osteoarthritis Patients Pubmed: 30526057
International Journal of Clinical and Experimental Medicine The relationship between the serum and synovial fluid osteopontin (OPN), cartilage oligomeric matrix protein (COMP), and the severity of knee osteoarthritis …
British Journal of Medicine and Medical Research Diagnostic Accuracy of Cartilage Oligomeric Matrix Protein (COMP), for Cartilage Damage in Rheumatoid Arthritis
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