ELISA Kit for Osteoclast Associated Receptor (OSCAR)

PIGR3; Osteoclast Associated,Immunoglobulin-Like Receptor; Polymeric immunoglobulin receptor 3; Poly-Ig receptor 3

Specificity

This assay has high sensitivity and excellent specificity for detection of Osteoclast Associated Receptor (OSCAR).
No significant cross-reactivity or interference between Osteoclast Associated Receptor (OSCAR) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Osteoclast Associated Receptor (OSCAR) and the recovery rates were calculated by comparing the measured value to the expected amount of Osteoclast Associated Receptor (OSCAR) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-97 94
EDTA plasma(n=5) 92-99 96
heparin plasma(n=5) 90-102 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Osteoclast Associated Receptor (OSCAR) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Osteoclast Associated Receptor (OSCAR) 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 Osteoclast Associated Receptor (OSCAR) 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) 87-95% 91-98% 89-97% 80-93%
EDTA plasma(n=5) 98-105% 81-104% 81-94% 87-97%
heparin plasma(n=5) 93-104% 89-96% 78-95% 95-104%

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
Journal of Molecular and Cellular Cardiology Quantitative proteomics reveals novel functions of osteoclast-associated receptor in STAT signaling and cell adhesion in human endothelial cells PubMed: 22985931
Arthritis Research & Therapy The immunoreceptor tyrosine-based activation motif (ITAM) -related factors are increased in synovial tissue and vasculature of rheumatoid arthritic joints PubMed: 23146195
Inflammation Osteoclast-Associated Receptor (OSCAR) Distribution in the Synovial Tissues of Patients with Active RA and TNF-α and RANKL Regulation of Expression by Osteoclasts In Vitro article:10.1007
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES Development of a novel polysaccharide-based iron oxide nanoparticle to prevent iron accumulation-related osteoporosis by scavenging reactive oxygen … Pubmed: 33049237
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