Description
ATP7B
All names and symbols: ATPase copper transporting beta; Copper-transporting ATPase 2; Copper pump 2; Wilson disease-associated protein; ATP7B; PWD; WC1; WD; WND
| Confirmed species reactivity | Human |
| Sensitivity | 0.056 ng/ml |
| Linear calculation range | 0.156 – 5 ng/ml |
| Precision intra-assay / inter-assay | <10% / <12% |
| Aeonian Rating® | 90 |
| RRID |
| Matrix | Recovery Range (%) | Average (%) |
| Serum (n=5) | 83-100 | 91 |
| EDTA plasma (n=5) | 91-100 | 95 |
| Heparin plasma (n=5) | 91-101 | 96 |
| Matrix | 1:2 | 1:4 | 1:8 | 1:16 |
| Serum (n=5) | 78-110% | 84-96% | 90-101% | 77-107% |
| EDTA plasma (n=5) | 93-96% | 89-109% | 72-106% | 90-95% |
| Heparin plasma (n=5) | 84-104% | 82-101% | 95-101% | 84-110% |
Calibratrion curve:
AEE0020 to ATP7B was successfully used to determine the concentration of ATP7B in serum, plasma and buffer. Recommended serum dilution 1:8, EDTA plasma dilution 1:2, and heparin plasma 1:8.
Calibration curve of recombinant ATP7B in buffer. HRP detection by Absorbance at 450nm.
Selectivity of the ELISA kit:
AEE0020 to ATP7B was successfully confirmed selective as there were no signals found in the presence of other closely related proteins at high and low concentrations.
Selectivity was tested by comparing the signals of recombinant ATP7B with other closely related recombinant proteins, each at two concentrations. The other related proteins tested were ATP7A, and ATOX1. HRP detection by Absorbance at 450nm.
ELISA Test:
The microtiter plate provided in this kit has been pre-coated with an antibody specific to ATP7B. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to ATP7B. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain ATP7B, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of ATP7B in the samples is then determined by comparing the O.D. of the samples to the standard curve.
ELISA Procedure:
1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 2 hours at 37℃;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37℃;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 1 hour at 37℃;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37℃;
8. Add 50µL Stop Solution. Read at 450nm immediately.
Stability:
The stability of ELISA 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 conditions.
Note:
To minimize unnecessary influences on the performance, operation procedures and lab conditions, especially room temperature, air humidity and incubator temperatures should be strictly regulated. It is also strongly suggested that the whole assay is performed by the same experimenter from the beginning to the end.
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