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Multiplex Assay Kit for Ferritin, Heavy Polypeptide (FTH) ,etc. by CBA (Cytometric Bead Array)

FTH1; FTHL6; PIG15; PLIF; Ferritin Heavy Chain; Apoferritin; Placenta Immunoregulatory Factor; Proliferation-Inducing Protein 15; Cell proliferation-inducing gene 15 protein

Specificity

This assay has high sensitivity and excellent specificity for detection of Ferritin, Heavy Polypeptide (FTH) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Ferritin, Heavy Polypeptide (FTH) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Ferritin, Heavy Polypeptide (FTH) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Ferritin, Heavy Polypeptide (FTH) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-103 96
EDTA plasma(n=5) 83-99 95
heparin plasma(n=5) 86-104 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Ferritin, Heavy Polypeptide (FTH) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Ferritin, Heavy Polypeptide (FTH) ,etc. by CBA (Cytometric Bead Array) 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 Ferritin, Heavy Polypeptide (FTH) ,etc. by CBA (Cytometric Bead Array) 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) 84-99% 85-98% 79-94% 88-102%
EDTA plasma(n=5) 89-96% 82-101% 98-105% 99-105%
heparin plasma(n=5) 96-104% 93-101% 79-102% 78-101%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
PLoS One Presence in the Pre-Surgical Fine-Needle Aspiration of Potential Thyroid Biomarkers Previously Identified in the Post-Surgical One PubMed: PMC3765839
Am J Physiol Lung Cell Mol Physiol. Methemoglobin-induced signaling and chemokine responses in human alveolar epithelial cells Pubmed:24142518
J Alzheimers Dis The Oral Iron Chelator, Deferasirox, Reverses the Age-Dependent Alterations in Iron and Amyloid-β Homeostasis in Rat Brain: Implications in the Therapy of Alzheimer’s Disease PubMed: 26484920
Eur Radiol MR molecular imaging of tumours using ferritin heavy chain reporter gene expression mediated by the hTERT promoter Pubmed:26960542
Exp Hematol. Impact of TET2 deficiency on iron metabolism in erythroblasts. pubmed:28167288
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