Multiplex Assay Kit for Hepcidin (Hepc) ,etc. by FLIA (Flow Luminescence Immunoassay)

HAMP; HFE2B; PLTR; LEAP1; Hepcidin Antimicrobial Peptide; Liver-expressed antimicrobial peptide 1; Putative liver tumor regressor

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

  • Multiplex Assay Kit for Hepcidin (Hepc) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Hepcidin (Hepc) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Specificity of the Multiplex Assay Kit for Hepcidin (Hepc) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

Recovery of the Multiplex Assay Kit for Hepcidin (Hepc) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 101
EDTA plasma(n=5) 80-93 81
heparin plasma(n=5) 91-99 95

Precision of the Multiplex Assay Kit for Hepcidin (Hepc) ,etc. by FLIA (Flow Luminescence Immunoassay)

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hepcidin (Hepc) ,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 Hepcidin (Hepc) ,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 of the Multiplex Assay Kit for Hepcidin (Hepc) ,etc. by FLIA (Flow Luminescence Immunoassay)

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Hepcidin (Hepc) ,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) 97-105% 98-105% 90-104% 95-102%
EDTA plasma(n=5) 97-104% 94-101% 80-95% 87-105%
heparin plasma(n=5) 86-104% 96-104% 89-103% 79-104%

Stability of the Multiplex Assay Kit for Hepcidin (Hepc) ,etc. by FLIA (Flow Luminescence Immunoassay)

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.

Assay procedure summary of the Multiplex Assay Kit for Hepcidin (Hepc) ,etc. by FLIA (Flow Luminescence Immunoassay)

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.

Test principle of the Multiplex Assay Kit for Hepcidin (Hepc) ,etc. by FLIA (Flow Luminescence Immunoassay)

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards,Labeled antigen and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest.A competitive inhibition reaction is launched between biotin labeled analytes of interest and unlabeled analytes of interest (Standards or samples) with the pre-coated antibody specific to analytes of interest. Following a wash to remove any unbound substances, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE) is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer. The MFI developed is reverse proportional to the concentration of analytes of interest in the sample.

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