ELISA Kit for Hemojuvelin (HJV)

HFE2; HFE2A; HJV; JH; RGMC; hemochromatosis type 2(Juvenile); Haemojuvelin; RGM domain family member C

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 102
EDTA plasma(n=5) 91-98 94
heparin plasma(n=5) 83-97 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hemojuvelin (HJV) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Hemojuvelin (HJV) 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 Hemojuvelin (HJV) 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) 80-103% 78-102% 78-97% 81-92%
EDTA plasma(n=5) 83-94% 92-102% 88-102% 94-102%
heparin plasma(n=5) 96-103% 79-94% 78-97% 91-103%

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 Citations
PLoS One. Effect of Iron Overload and Iron Deficiency on Liver Hemojuvelin Protein PubMed: PMC3356351
Journal of Hepcidin Levels Are Increased in Patients with Acute Ischemic Stroke: Preliminary Report PubMed: 25881778
European Journal of Haematology Soluble hemojuvelin in transfused and untransfused thalassaemic subjects doi:10.1111
American Journal of Nephrology The secret war against counterfeit science pubmed:28492262
Critical Care Urinary biomarkers predict advanced acute kidney injury after cardiovascular surgery Pubmed:29699579
Scientific Reports Hemojuvelin Predicts Acute Kidney Injury and Poor Outcomes Following Cardiac Surgery Pubmed:29386545
Journal of Cerebral Blood Flow and Metabolism The functional role of hemojuvelin in acute ischemic stroke Pubmed: 31307288
Oxidative damage and iron dyshomeostasis in thalassemic patients
J Trace Elem Med Biol Caffeine and alcohol-friends or foes of human iron stores? Pubmed:35063815
Arch Pathol Lab Med Urinary Biomarkers Can Predict Weaning From Acute Dialysis Therapy in Critically Ill Patients Pubmed:35311928
Arch Pathol Lab Med The Relationships among Plasma Fetuin-B, Thyroid Autoimmunity, and Fertilization Rate In Vitro Fertilization and Embryo Transfer Pubmed:35340679
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