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小鼠血管紧张素转化酶(ACE)检测试剂盒
ELISA Kit for Angiotensin I Converting Enzyme (ACE)
| Organism species | Mus musculus (Mouse) |
| Product No. | YBA004Mu |
| Sample type | Serum, plasma, tissue homogenates, cell lysates, cerebrospinal fluid, cell culture supernates and other biological fluids |
| Format | 96T |
| Assay length | 4.5 hours |
| Detection range | 0.313-20ng/mL The standard curve concentrations used for the ELISA’s were 20ng/mL, 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.313ng/mL |
| Sensitivity | The minimum detectable dose of this kit is typically less than 0.135ng/mL. |
No significant cross-reactivity or interference between Angiotensin I Converting Enzyme (ACE) and analogues was observed.
小鼠血管紧张素转化酶(ACE)检测试剂盒
Recovery
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 98-105 | 102 |
| EDTA plasma(n=5) | 96-104 | 99 |
| heparin plasma(n=5) | 98-105 | 101 |
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Angiotensin I Converting Enzyme (ACE) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
小鼠血管紧张素转化酶(ACE)检测试剂盒
Linearity
| Sample | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 84-99% | 81-103% | 90-99% | 87-104% |
| EDTA plasma(n=5) | 89-101% | 78-102% | 81-96% | 79-102% |
| heparin plasma(n=5) | 82-105% | 79-89% | 80-102% | 78-99% |
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 | Quantity | Reagents | Quantity |
| Pre-coated, ready to use 96-well strip plate | 1 | Plate sealer for 96 wells | 4 |
| Standard | 2 | Standard Diluent | 1×20mL |
| Detection Reagent A | 1×120µL | Assay Diluent A | 1×12mL |
| Detection Reagent B | 1×120µL | Assay Diluent B | 1×12mL |
| TMB Substrate | 1×9mL | Stop Solution | 1×6mL |
| Wash Buffer (30 × concentrate) | 1×20mL | Instruction manual | 1 |
小鼠血管紧张素转化酶(ACE)检测试剂盒
Assay procedure summary
2. Add 100µL standard or sample to each well. Incubate 2 hours at 37oC;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37oC;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37oC;
8. Add 50µL Stop Solution. Read at 450nm immediately.
小鼠血管紧张素转化酶(ACE)检测试剂盒
Test principle
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Angiotensin I Converting Enzyme (ACE). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Angiotensin I Converting Enzyme (ACE). 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 Angiotensin I Converting Enzyme (ACE), 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 Angiotensin I Converting Enzyme (ACE) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
