Multiplex Assay Kit for Procollagen III C-Terminal Propeptide (PIIICP) ,etc. by FLIA (Flow Luminescence Immunoassay)

P3CP; C-Propeptide Of Type III Procollagen; Procollagen III Carboxy Terminal Propeptide

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Procollagen III C-Terminal Propeptide (PIIICP) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Procollagen III C-Terminal Propeptide (PIIICP) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Procollagen III C-Terminal Propeptide (PIIICP) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Procollagen III C-Terminal Propeptide (PIIICP) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-94 88
EDTA plasma(n=5) 93-102 97
heparin plasma(n=5) 81-91 85
sodium citrate plasma(n=5) 91-98 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Procollagen III C-Terminal Propeptide (PIIICP) ,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 Procollagen III C-Terminal Propeptide (PIIICP) ,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 Procollagen III C-Terminal Propeptide (PIIICP) ,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) 79-101% 79-98% 92-101% 91-99%
EDTA plasma(n=5) 97-105% 94-102% 98-105% 84-96%
heparin plasma(n=5) 92-101% 82-90% 81-92% 97-104%
sodium citrate plasma(n=5) 79-96% 93-101% 89-98% 82-101%

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#:PIIICP) 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 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
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Heart & Vessels Left ventricular reverse remodeling is not related to biopsy-detected extracellular matrix fibrosis and serum markers of fibrosis in dilated cardiomyopathy, regardless of the definition used for LVRR pubmed:28004175
MOLECULAR MEDICINE REPORTS Endothelial‑to‑mesenchymal transition in human idiopathic dilated cardiomyopathy pubmed:29115553
Heart and Vessels Left ventricular reverse remodeling is not related to biopsy‑detected extracellular matrix fibrosis and serum markers of fibrosis in dilated cardiomyopathy, regardless of the definition used for LVRR 10.1007/s00380-016-0930-y
Cytokine 12-month patterns of serum markers of collagen synthesis, transforming growth factor and connective tissue growth factor are similar in new-onset and chronic dilated cardiomyopathy in patients both with and without cardiac fibrosis. pubmed:28460256
Advanced Healthcare Materials Prognostic value of fibrosis-related markers in dilated cardiomyopathy: A link between osteopontin and cardiovascular events. pubmed:29120858
JOURNAL OF THROMBOSIS AND THROMBOLYSIS Extracellular matrix collagen biomarkers levels in patients with chronic thromboembolic pulmonary hypertension Pubmed: 33175289
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