Multiplex Assay Kit for Alpha-Endorphin (aEP), 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.

1. Preparation of standards, reagents and samples before the experiment; <br/> 2. Add 50μL standard or sample to each well, <br/> &nbsp; &nbsp; &nbsp; &nbsp; add 10μL magnetic beads, and 50μL Detection Reagent A, incubate 90min at 37&deg; C on shaker; <br/> 3. Wash plate on magnetic frame for three times; <br/> 4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37&deg; C on shaker; <br/> 5. Wash plate on magnetic frame for three times; <br/> 6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.<br/>

<p> Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Alpha-Endorphin (aEP), etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively. <br/> Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Alpha-Endorphin (aEP), etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate. <br/> CV (%) = SD/meanX100 <br/> Intra-Assay: CV<10% <br/> Inter-Assay: CV<12% <br/> </p>

<p> 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. <br/> 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. </p>

<p> This assay has high sensitivity and excellent specificity for detection of Alpha-Endorphin (aEP), etc. by FLIA (Flow Luminescence Immunoassay) .<br/> No significant cross-reactivity or interference between Alpha-Endorphin (aEP), etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed. </p>