This assay has high sensitivity and excellent specificity for detection of N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP). No significant cross-reactivity or interference between N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) and analogues was observed.
精密度:
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100; Intra-Assay: CV<10%; Inter-Assay: CV<12%.
稳定性:
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.
检测原理:
The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP). A competitive inhibition reaction is launched between biotin labeled N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) and unlabeled N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) (Standards or samples) with the pre-coated antibody specific to N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP). 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 N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) in the sample. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is reverse proportional to the N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) level in the sample or standard.
实验流程:
1. Prepare all reagents, samples and standards;
2. Add 50ul standard or sample to each well.
And then add 50ul prepared Detection Reagent A immediately.
Shake and mix. Incubate 1 hour at 37℃;
3. Aspirate and wash 3 times;
4. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37℃;
5. Aspirate and wash 5 times;
6. Add 100ul Substrate Solution. Incubate 10 minutes at 37℃;
7. Read RLU value immediately.