Multiplex Assay Kit for Orexin A (OXA) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Orexin A (OXA) ,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 Orexin A (OXA) ,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%

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Orexin A (OXA) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

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.

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.