Multiplex Assay Kit for Beta-Crosslaps (bCTx) ,etc. by FLIA (Flow Luminescence Immunoassay)

b-CTx; bCTXI; bCTX-I; B-Cr; BCL; Type I Collagen C-Telopeptide-Related Fraction

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Beta-Crosslaps (bCTx) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Beta-Crosslaps (bCTx) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 84-103 88
EDTA plasma(n=5) 80-103 81
heparin plasma(n=5) 92-101 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Beta-Crosslaps (bCTx) ,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 Beta-Crosslaps (bCTx) ,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 Beta-Crosslaps (bCTx) ,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) 80-92% 87-97% 80-96% 89-101%
EDTA plasma(n=5) 80-99% 90-99% 90-104% 83-92%
heparin plasma(n=5) 81-93% 78-89% 87-96% 81-101%

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#:bCTx) 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 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Bosnian Journal of Basic Medical Sciences Effects of carbamazepine on serum parathormone, 25-hydroxyvitamin D, bone specific alkaline phosphatase, C-telopeptide, and osteocalcin levels in healthy rats PubMed: 23198939
Clin Cancer Res. Cabozantinib inhibits prostate cancer growth and prevents tumor-induced bone lesions Pubmed:Pmc3946460
Bone Elevated serum soluble CD200 and CD200R as surrogate markers of bone loss under bed rest conditions Pubmed:24333170
J Bone Miner Metab. Effect of Urocortin on strength and microarchitecture of osteopenic rat femur Pubmed:24633537
Journal of Nutrition Cholecalciferol Supplementation Promotes Bone Turnover in Chinese Adults with Vitamin D Deficiency Pubmed:29897564
Journal of Functional Foods Effect of long-term administration of mangiferin from on bone metabolism in ovariectomized rats 10.1016:j.jff.2018.04.048
European Journal of Medical Research Biomechanical and histological analyses of the fracture healing process after direct or prolonged reduction Pubmed: 30180907
Endokrynologia Polska FGF23 and primary hyperparathyroidism: is there a link? Pubmed: 32598019
血清骨膜素水平与老年髋部骨折患者骨密度相关性研究.
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