ELISA Kit for Procollagen III C-Terminal Propeptide (PIIICP)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 94-104 98
EDTA plasma(n=5) 79-99 92
heparin 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) 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) 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) 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) 84-98% 78-96% 83-95% 78-96%
EDTA plasma(n=5) 80-96% 81-95% 78-101% 79-90%
heparin plasma(n=5) 80-93% 81-91% 86-101% 95-105%

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 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Inflammation Research Fibrosis of extracellular matrix is related to the duration of the disease but is unrelated to the dynamics of collagen metabolism in dilated cardiomyopathy pubmed:27516211
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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