Low Sample Volume ELISA Kit for Alpha-Endorphin (aEP)

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Mini Samples Alpha-Endorphin (aEP) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Mini Samples Alpha-Endorphin (aEP) and unlabeled Mini Samples Alpha-Endorphin (aEP) (Standards or samples) with the pre-coated antibody specific to Mini Samples Alpha-Endorphin (aEP) . After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Mini Samples Alpha-Endorphin (aEP) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Mini Samples Alpha-Endorphin (aEP) in the sample.

1. Prepare all reagents, samples and standards; <br/> 2. Add 25&micro; L standard or sample to each well.<br/> &nbsp; &nbsp; &nbsp; &nbsp; And then add 25μL prepared Detection Reagent A immediately.<br/> &nbsp; &nbsp; &nbsp; &nbsp; Shake and mix. Incubate 1 hour at 37&deg; C; <br/> 3. Aspirate and wash 3 times; <br/> 4. Add 50&micro; L prepared Detection Reagent B. Incubate 30 minutes at 37&deg; C; <br/> 5. Aspirate and wash 5 times; <br/> 6. Add 50&micro; L Substrate Solution. Incubate 10-20 minutes at 37&deg; C; <br/> 7. Add 25&micro; L Stop Solution. Read at 450 nm immediately.<br/>

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Mini Samples Alpha-Endorphin (aEP) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Mini Samples Alpha-Endorphin (aEP) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV Inter-Assay: CV Inter-Assay: CV

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.

This assay has high sensitivity and excellent specificity for detection of Mini Samples Alpha-Endorphin (aEP) . No significant cross-reactivity or interference between Mini Samples Alpha-Endorphin (aEP) and analogues was observed.