Multiplex Assay Kit for Hemojuvelin (HJV) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 88-98 94
EDTA plasma(n=5) 80-97 87
heparin plasma(n=5) 95-103 98
sodium citrate plasma(n=5) 86-101 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hemojuvelin (HJV) ,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 Hemojuvelin (HJV) ,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 Hemojuvelin (HJV) ,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) 96-105% 79-99% 78-104% 81-98%
EDTA plasma(n=5) 90-98% 93-102% 84-96% 87-102%
heparin plasma(n=5) 86-95% 89-99% 86-102% 90-101%
sodium citrate plasma(n=5) 98-105% 98-105% 90-101% 89-101%

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#:HJV) 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 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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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