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ELISA Kit for Ferritin (FE)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 79-97 90
EDTA plasma(n=5) 83-99 93
heparin plasma(n=5) 83-102 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Ferritin (FE) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Ferritin (FE) 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 (FE) 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) 78-96% 96-103% 91-103% 87-99%
EDTA plasma(n=5) 89-102% 78-88% 93-101% 79-93%
heparin plasma(n=5) 90-101% 94-105% 88-96% 97-104%

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 Reference
Molecular Endocrinology PGC-1α regulates hepatic hepcidin expression and iron homeostasis in response to inflammation. Pubmed: 23438894
experimental oncology Redox-regulation of gelatinases during growth of cisplatin-sensitive and resistant Guerin carcinoma. PubMed: 25804229
J Cancer Sci Ther Host H68D Genotype Affects Tumor Growth in Mouse Melanoma Open-Access: 1948-5956-1000353
Food Funct The effects of polysaccharides from the root of Angelica sinensis on tumor growth and iron metabolism in H22-bearing mice Pubmed:26757699
Physiological research The Labile Iron Pool in Monocytes Reflects the Activity of the Atherosclerotic Process in Men with Chronic Cardiovascular Disease pubmed:27782743
Expert Review of Hematology 4 (th) Mediterranean Multidisciplinary Course on Iron Anemia April 29 (th)-30 (th) 2016, Madrid, Spain. pubmed:27615544
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