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OneStep ELISA Kit for Aspartate Beta Hydroxylase (ASPH)

BAH; CASQ2BP1; HAAH; JCTN; Junctin; Humbug; Junctate; Aspartyl/Asparaginyl Beta-Hydroxylase; Peptide-aspartate beta-dioxygenase

Specificity

This assay has high sensitivity and excellent specificity for detection of Aspartate Beta Hydroxylase (ASPH).
No significant cross-reactivity or interference between Aspartate Beta Hydroxylase (ASPH) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Aspartate Beta Hydroxylase (ASPH) and the recovery rates were calculated by comparing the measured value to the expected amount of Aspartate Beta Hydroxylase (ASPH) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-102 98
EDTA plasma(n=5) 85-96 90
heparin plasma(n=5) 78-92 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Aspartate Beta Hydroxylase (ASPH) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Aspartate Beta Hydroxylase (ASPH) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Aspartate Beta Hydroxylase (ASPH) 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) 99-105% 85-99% 92-101% 82-105%
EDTA plasma(n=5) 99-105% 85-93% 86-101% 99-105%
heparin plasma(n=5) 95-103% 85-96% 88-95% 78-96%

Stability

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.

Reagents and materials provided

Reagents Quantity Reagents Quantity

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well, then add 50µL prepared Detection Reagent A to each well. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
5. Add 50µL Stop Solution. Read at 450nm immediately.

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